Plastic Degradation by Extremophilic Bacteria
Round 1
Reviewer 1 Report
Comments included in the attached file.
Comments for author File: 
Comments.pdf
Author Response
All cited references need to be checked carefully, both in text and in the Bibliography. This is for 2 reasons:
- In the manuscript as submitted there is a discrepancy between the citation
. number in the text and the corresponding number in the Bibliography. The reason
. for this appears to be:
Text. Bibliography
[1]……corresponds to…….[1]
one of these references in the Bibliography
Is not included in the text, and as a consequence [9]……corresponds to……[10]
all in this sequence are
therefore -1 compared
to the Bibliography
[25]…..corresponds to……[26]
[27] this reference (Pinto et al) is not included in the text, and as a consequence
[26]…..corresponds to……[28]
all in this sequence are
therefore -2 compared
to the Bibliography
[62]…..corresponds to……[64]
- The revised text has suggested the introduction of new additional references
. which will consequently require a revision of the numbering of the other
. subsequent references both in text and in the Bibliography.
Sorry for the discrepancy in the numbering of references. This version was carefully checked and hope now numbering is correct.
Page 1 Line 36: human life. Versatility ….
Replace with: human activity. Versatility ……
The word “life” was replaced by “activity”
Page 1 Line 39: production cost, easy manufacturing …… Replace with: production cost, ease of manufacturing …..
“Ëasy”was changed by “ease of”
Page 1 Line 45: rate in 2020: -8.5%. Replace with: rate in 2020 of -8.5%
“of” was added
Page 2 Line 5: 1, plastic producers are spread worldwide with a biggest deal of Asia
Replace with: 1, plastic producers are spread worldwide, the biggest contributors
. being Asia
The phrase was changed according to the suggested way
Page 2 Line 16: based wrapping materials. Their packaging application accounts for
. around 40% of the plastic
Replace with: based wrapping materials , and now account for around 40% of the.
. plastics produced.
The phrase was changed according to the suggested way
Page 5 Line 13: a slow process [15]. That’s why …..
Replace with: a slow process [15]. That is why …..
“That’s” was replaced by “That is”
Page5 Lines 23
+ 24: The resistance of plastics against microbial attack is a result of a
. short evolutionary time of their presence in nature that is not
. enough for design of
Replace with: The resistance of plastics to microbial attack results from the
. short time of their presence in nature not being enough for the
. evolution of
Replaced
Page 5 Line 26: Enzymes involved in plastic degradation processes belong to the
. groups
Replace with: Characterised enzymes known to be involved in plastic degradation
. processes comprise a relatively small group
This part was changed according to the advice of another reviewer.
Page 5 Lines29
+ 30: ………………………… active mainly on aliphatic polyesters. Bacterial
polyurethane esterases have been reported to degrade polyurethanes
Replace with: …………………………..active mainly on aliphatic polyesters. Examples
. of bacterial esterases able to degrade polyurethanes have been
. reported [19].
Replaced
Page 6 Lines 1
+ 2: Cutinase (EC 3.1.1.74) is known as a hydrolase for a variety of
. polymers although initially has been described as able to ….
Replace with: Cutinase (EC 3.1.1.74) is a known hydrolase for a variety of
. polymers although initially characterised as able to ….
The sentence was changed.
Page 6 Line 6: An new additional reference is needed for any examples of proteases . able to hydrolyse polyamide-type synthetic polymers such as nylons - . otherwise it needs to be made clear that while proteases can hydrolyse
. natural polyamides (such as proteins), this ability is yet to be confirmed
. with synthetic polyamides (as is implied by the text included in Page 6
. lines 8-10 incl).
The reference Tomita et al., 2003 was included
Page 6 Line 8: Still the information …..
Replace with: Currently, the information ……
Replaced
Page 6 Line 10
+ 11: ……….[20]. The reports for their degradation
. concerns mainly polymer …………..
Replace with: ……….[20]. The reports of their degradation
. concern mainly polymer …………..
Replaced
Page 6 Line13: responsible for their accumulation. Although a degradation of .. Replace with: responsible. Although some degradation of …………
Replaced
Page 6 Lines16
+ 17: degradation but to the attack to the chemical additives in their
. molecules [17]. As an example polystyrene sultanate …
Replace with: degradation , but to the metabolism of the relevant chemical
. adjunct molecules [17]. Similarly, polystyrene sultanate …
Replaced
Page 6 Lines 20
+ Line 21: Seeking for new plastic active enzymes and microorganisms .
. determines the interest toward extremophiic enzymes due to the
. change…………..
Replace with: The search for new plastic active enzymes and microorganisms .
. has resulted in interest in relevant extremophiic enzymes due to the
. changes that occur
Replaced
Page 6 Line 23
+ 24 …… water content and nutrients, high radiation. Such enzymes are
. produced by extremophilic microorganisms
Replace with: ……. content of water and other nutrients, and high radiation. It is
. known that extremophilic microorganisms are competent producers .
of a range of potentially relevant hydrolytic enzymes [ * ]
* a new relevant reference is required here eg ‘Extremophiles for
. hydrolytic enzyme production: biodiversity and potential
. biotechnological applications’ D. Kour et al.,
. doi.org/10.1002/9781119434431.ch16
The sentence “Such enzymes…” was removed.
Page 6 Line 26: ….. methods for registration of microbial
Replace with: ….. methods for establishing relevant microbial
This phrase was changed
Page 6 Line 28: ……. changes in plastic like appearance of holes or
Replace with: ……. changes in plastic such as appearance of holes or
Replaced
Page 6 Line 31: ………………a good approach for sophisticated
Replace with: ………………a good approach for more sophisticated
Replaced
Page 6 Line 42: 5. Registration of products ………….
Replace with: 5. Detection and characterisation of products ………..
Replaced
Page 7 Figure 4: hallo Replace with: halo
Page 7 Lines 8
+ 10: ….………….…….the substrate. The biologically relevant structure
. appropriate for such a purpose is the biofilm [23]. Biofilms represent
. consortia of microorganisms attached to surfaces and embedded in
.Replace with: ….………….…….the substrate. Biofilms are biologically relevant
. structures appropriate for such a purpose[ 23]. Biofilms are
. consortia of microorganisms attached to and/or embedded in an .
Replaced
Page 7 Lines17 ………rate of macromolecules, including
+ 18: enzymes, through the biofilm matrix, are prerequisites for
Replace with:: ………rate of macromolecules, including extracellular
. enzymes, through the biofilm matrix, are all factors that favour
Replaced
Page 8 Lines 1
+ 2: ……. conditions are significant factors
. [26], the nature of the colonising organisms is decisive.
Replace with: ……. conditions are both significant abiotic factors
[26], while the nature of the colonising organisms is clearly decisive.
Replaced
Page 8 Lines 8
+ 14: ….. degradation. For example, Klebsiella pneumonia CH001
. [27] and Rhodococcus sp. [24] formed biofilm which was related with
. degradation of PE, biofilms by Pseudomonas citronellolis and Bacillus flexus
. showed degradation activity towards PVC [28], B. subtilis ET18 and B. .
- cereus ET30 formed biofilms on nylon and PET causing damage of the
. plastic surface [29]. Combinations of two species of bacteria were
. shown to enhance the plastic-biodegradation potential. Lately, the attention has
- Replace with: . ….. degradation. For example, single species biofilms of
. Klebsiella pneumonia CH001 [27] and Rhodococcus sp. [24] promoted
. the degradation of PE, and corresponding biofilms of Pseudomonas
. citronellolis and Bacillus flexus showed degradation activity towards
. PVC [ * ] ; similarly, B. subtilis ET18 and B. cereus ET30 each formed
. single species biofilms on nylon and PET causing damage to the
. plastic surface [29]. Combinations of two species of bacteria were shown .
. to enhance the plastic-biodegradation potential [ ** ]. Lately, the attention has
Replaced
. [ * ]. Reference [28] currently shown in text actually refers to original
. research reported in the review currently listed as [29] in the
. Bibliography. The correct reference to the relevant original scientific
. paper is: Giacomucci L. et al., New Biotechnol. 52, 35 - 41 (2019)n.
.
[ ** ] There is currently no relevant reference cited in text, but this is
. required. A suitable paper for this purpose would be Pinto, M. et al
. PLoS ONE 2019, 14. - currently listed as [27] in the Bibliography.
Relevant bibliography is listed
Page 8 Lines 17
+ 18: ………… formation and biodegradation, to
. the present moment the molecular mechanism ….
- Replace with: ………… formation and biodegradation,
. currently the molecular mechanism ….
Changed
Page 8 Line 20: …….and enzymatic activities originating from biofilm
Replace with: …….and enzymatic activities specifically originating from
biofilm
Replaced
Page 8 Line 23: Mostly extremophiles belong to Bacteria and Archaea.
Replace with: Most currently known extremophiles are either Eubacteria or Archaea.
Replaced
Page 8.Lines 25 extremophilicity is usually significantly lower and consequently could not
+ 26: influence the plastic properties to such an extent as prokaryotes do.
- Replace with: extremophilicity is usually significantly lower and consequently they
. have less potential than extremophilic prokaryotes to influence the
. properties of plastics.
Replaced
Page 8 Line 29: …..archaeal degraders is not yet reported.
Replace with: …..archaeal degraders has not yet been reported.
Replaced
Page 8 Line 30: That’s why extremophilic eucaryotes …..
Replace with: For these reasons extremoophilic eucaryotes …..
Replaced
Page 8 Lines 33
+ 34: …..to grow in lethal or too harsh for human
- existence conditions. It is generally ………..
- Replace with: …..to grow in conditions that would be lethal or too
. harsh for human existence. It is generally ………..
Replaced
Page 8 Lines 36
- 38: species available in natural ecosystems and in the extreme niches this
. amount is especially low due to difficulties to reproduce specific
.. environmental conditions in the laboratory. Microbes from extreme
- Replace with: species present in natural ecosystems : this fraction is especially low.
. in samples taken extreme niches due to difficulties in reproducing
. such environmental conditions in the laboratory Consequently,
- . microbes from extreme
- Replaced
- Page 8 Lines 40 ……..industrial applications. Among
- - 42: biomolecules the extremophilic enzymes (extremozymes) are of .
special interest for biotechnology, biomedicine and industrial .
processes due to
- Replace with: ….. industrial applications. Of special
. interest for biotechnology, biomedicine and industrial processes are
. the enzymes (extremozymes) produced by extremophiles due to
Replaced
-
Page 8 Line 45: producers represent a challenge for resolving ……….
Replace with: producers may hold the potential for resolving ……….
Replaced
Page 8 Lines 49
- 52: Thermophiles are heat loving microorganisms that need high
- temperatures for their growth between 45 and 122oC [30]. Thermophiles
. literally heat lovers are organisms that grow at temperatures above those
. that sustain most life forms. According to their growth temperature
- Rep[lace with: Thermophiles, literally heat lovers, are microorganisms that need high
- temperatures (between 45 and 122oC) for their growth [30] : as such,
. they are organisms that grow at temperatures above those that sustain
. most life forms. According to their growth temperature range,
Replaced
Page 9 Line 1: thermophiles are classified in several groups, facultative thermophiles Replace with: they are classified in several groups ; facultative thermophiles
This paragraph was rewritten.
Page 9 Line 5: 80oC. Growth of hyperthermophiles ….
Replace with: 80oC , while growth of hyperthermphiles …….
This paragraph was rewritten.
Page 9 Line 7: ecosystems such as deep-sea and terrestrial hot springs, geysers,
Replace with: ecosystems such as deep-sea black smokers, terrestrial hot springs and
. geysers,
Replaced
Page 9. Lines 12
- 14: proteins, shorter protein length, participation of heat shock proteins in
. protein folding; stabilisation of membranes by branched chain fatty acids
. and polyamides; active system for repairing of DNA damages [32].
Replace with: proteins, shorter protein length, and the participation of heat shock
. proteins in protein folding; additionally they have evolved more stable
. membranes by incorporating branched chain fatty acids and
. polyamides , and active systems for repairing DNA damage [32].
Replaced
Page 9 Line 15: …..They are divided in
Replace with: …..They are divided into
Replaced
Page 9 Line 18: They have developed systems for influx of …….
Replace with: They have developed systems to regulate the influx of …….
Replaced
Page 9 Lines 20
+ 21: …… alkaline soda lakes,
Replace with: man-made environments as a result of agricultural activity.
- … alkaline soda lakes, and some
- man-made environments that result from agricultural activity.
Replaced
-
Page 9 Line 24: …..growth by an increased …
Replace with: …..growth by a number of different strategies including an increased…
Replaced
Page 9 Line 26: ……… membrane fluidity ; high synthesis ….. Replace with: ……… membrane fluidity , high synthesis …..
Replaced
Page 9 Lines 27
- 29: proteins ; synthesis of anti-freeze proteins that bind to ice crystals ; .
accumulation of compatible solutes as cry-protectants to prevent cell .
damage ; adaptation of psychrophlic enzymes activity to low temperature
- Replace with: proteins , synthesis of anti-freeze proteins that bind to ice crystals ,
. accumulation of compatible solutes as cry-protectants to prevent cell .
damage , and adaptation of psychrophlic enzymes to activity at low
Replaced
-
Page 9 Line 39: strategies , maintaining more water ………
Replace with: strategies; either maintaining more water ………
Replaced
Page 9 Line 44: Evolving in harsh conditions …..
Replace with: As a general principle, evolving in harsh conditions …..
Replaced
Page 9 Line 46: solvents and detergents. Attractive feature of …….
Replace with: solvents and detergents. An attractive feature of …….
Replaced
Page 9 Line 47: enzymes is slowing down of ………
Replace with: enzymes is the slowing down of ………
Replaced
Page 9 Line 49: ….. Their long life prevents a …..
Replace with: ….. Their long life is beneficial because it potentially prevents a …..
Replaced
Page 9 Line 50: ……………..like plastic degradation. Many plastic-
Replace with: ……………..like plastic degradation. Furthermore, many plastic-
Replaced
Page 9 Line 53: ……or high pressure. Analysis of ……
Replace with: ……or high pressure. However, analysis of ……
The whole sentence was changed
Page 10. Lines 2
- 7: …… moderate extremophiles . The reason for
. very limited information for degradation by extremophiles living in very
. extreme conditions could be short time for evolving metabolic .
mechanisms of extremophilic ………………..difficult substrates ; .
the enhanced sensitivity. ……..………………………… in the case of .
thermophiles, low growth rate ……….
- Replace with: …… moderate extremophiles ,whereas there is . The reason for . very limited information for degradation by extremophiles living in very
. extreme conditions. Possible reasons for this discrepancy, which are relevant
. to all categories of extremophiles, could be the short time for evolving metabolic
Replaced
- mechanisms of extremophilic ……………………difficult substrates , allied to
. enhanced sensitivity ……..……………………………………. in the case of
. thermophiles, and the low growth rate ……….
Corrected
Page 10 Line11: ………………….thermal habitats suggests an improved
Replace with: …..thermal habitats is potentially advantageous because of improved
Replaced
Page 10 Line15: are the higher enzyme turnover rate as a …….. Replace with: are the higher rates of enzyme activity as a ……
Replaced
Page 11. Table 1: This Table needs to be reformatted. Where it is not possible to include
. whole words, they need to be hyphenated in a ‘sensible’ way, eg
Degrad- Poly- Biodegrad- Polycapro-
ation ethylene able lactone
In addition, Mw needs to be amended to MW
Changed
Page 12 Line 7: degradation by a thermophilic actinomycete …….
Replace with: degradation by a pure culture of one of the microorganisms, the
. thermophilic actinomycete ……..
Replaced
Page 12 Line 8: analysed by PDS-GPC …………. Replace with: when analysed by PDS-GPC …….
Replaced
Page 12 Line 9: …………molecular weight of the used PLC began to
- 22 decrease gradually at 72 h of its cultivation accompanied by appearance
of oligomeric peaks. Further degradation of oligomers was demonstrated
by lowering of their peaks suggesting that this strain could achieve full
mineralization of PCL and decomposed 38% of plastic after 6 days of
composting. However, when Bacillus licheniformis HA1 isolated from the
same compost was added a significant increase in PCL degradation was
observed reaching a value of 70% after 48 h. B. licheniformis HA 1 alone
was not able to utilise plastic, however it grew by degradation of the
intermediates and adjusted pH. Synergistic effect in the simultaneous
cultivation of both thermophilic strains accelerated PCL degradation and
. increased significantly the portion of the decomposed polymer. The
constant concentration of the primary degrading microorganism was
suggested to be a result of the low rate of plastic degradation as a rule.
Replace with: …………molecular weight of the initial PLC polymer began to
. decrease gradually 72 h after cultivation commenced, accompanied
by the appearance of oligomeric peaks. Further degradation of the resultant oligomers was demonstrated by the lowering of the relevant peaks suggesting that this strain could
achieve extensive mineralization of PCL , resulting in a 35% decomposition of the plastic
after 6 days of composting. However, when Bacillus licheniformis HA1 , the synergistic
partner isolated from the same compost was added , a significant increase in PCL
degradation was observed reaching a value of 70% after 48 h. It was suggested that
while B. llcheniformis HA 1 alone was not able to utilise plastic, it was able to grow by
degradation of the intermediates released by S. thermodenitrificans and the consequent
altered pH. The synergistic effect resulting from the simultaneous cultivation of both
thermophilic strains accelerated PCL degradation and increased significantly the portion of the decomposed polymer. It was suggested that the constant concentration of the primary degrading microorganism was a result of the low rate of plastic degradation.
Replaced
Page12 Line 25: Two PCL-degrading enzymes ……..
Replace with: Two PCL-degrading extracellular enzymes ……..
Replaced
Page 12 Line28: ……[42]. Reduction of gravimetric and
Replace with: ……[42]. In a similar report a reduction of the gravimetric and
Replaced
Page 12 Line 32: was a poor biofilm-former [35]. A thermophilic isolate ………
Replace with: was a poor biofilm-former [35]. Although less well characterised, a
. thermophilic isolate ……
Replaced
Page 12 Line 43: Thermomonospora, species Thermobifida alba [45], …..
Replace with: Thermomonospora, specifically the species Thermobifida alba [45], …
Replaced
Page 12 Line 45 ………………after 3 weeks action of the
- 49: polyester hydrolyse TfH synthesised from a thermophilic bacterium
. Thermobifida fascia [18]. The recombinant T. fusca cutinase TfCut2 was over
. expressed in B.subtilis as the dominant protein in the supernatant
. after 42 h of cultivation at 37oC which was able to degrade lcPET film
- Replace with: ……………after 3 weeks of action by the
. extracellular polyester hydrolyse TfH secreted by the thermophilic bacterium
. Thermobifida fascia [ * ] When the recombinant T. fusca cutinase TfCut2 was over
. expressed in B.subtilis, it was the dominant protein in the supernatant , which
. after 42 h of cultivation at 37oC was able to degrade lcPET film
v Replaced
[ * ] Correct reference citation required. - unlikely to be [18]
Corrected
Page 13 Line13: Nylon is a synthetic polyamide characterised ……..
Replace with: Nylon is the generic name for a related group of synthetic polyamides
. characterised ……..
Replaced
Page13 Line15: … intermolecular hydrogen bonds between molecular chains ……. Replace with: … intermolecular hydrogen bonds between the polymeric chains …….
Replaced
Page 13 Line 16 numerical nomenclature depends from the ratio between the number of
- 18: carbon atoms in the diamine and dibasic acid monomers used for their
. manufacturing.
Replace with; numerical nomenclature depends on the number of
. carbon atoms in the monomers used for their
. manufacture.
Replaced
Page 13 Lines 20
+ 21: ………………. nylon 12 and 6 at 60oC,
. however was not able to degrade nylon 66, more crystalline than other two nylons.
- Replace with: ………………. nylon 12 and 6 at 60oC,
. but was not able to degrade the more crystalline polymer nylon 66 [39].
Replaced
Page13 Line 23: The reported thermophilic …….
Replace with: All the above reported thermophilic ……….
Replaced
Page13 Lines 24 ……obligate thermophiles and to the best of our knowledge
- 27: extreme and hyperthermophiles are not known to the best of our
- knowledge. Similarly to mesophiles thermophilic Bacillaceae members
. are a good source of enzymes for plastic transformation bioprocesses.
- Replace with: ……obligate thermophiles, with thermophilic Bacillaceae being a
. good source of enzymes for plastic transformation bioprocesses . and to the best of
our knowledge equivalent competent extreme and hyperthermophiles are currently . not known.
Replaced
Page 13 Lines 29 pH is another environmental factor the affects solubility and
- 31: softening of plastics. To the best of our knowledge reports for a capability
. to degrade synthetic polymers by acidophiles (excepting fungi) are not
. available and alkaliphiles are scarcely investigated. …..
Replace with: pH is another environmental factor that can affect both the solubility and
. softening of plastics. However, the ability to degrade synthetic
. polymers by acidophilic bacteria has been scarcely investigated. …..
v Replaced
Page 13 Lines 33
- 35: ……. hyperalkaline waters (pH 11)
. from the spring Poon Bate, Betelan, Zambales, Philippines [51]. Nine
. strains were isolated after enrichment at a synthetic medium
. supplemented with ldPE as a sole source of carbon pH 11
Replace with: ……. hyperalkaline water samples (pH 11)
. from a spring in the Philippines [51]. Nine strains were isolated after
. enrichment in a synthetic medium supplemented with ldPE as a sole
. source of carbon and adjusted to pH 11
Replaced
Page 13 Lines 38
- 40: …… and a Bacillus sp. Each of them separately reduced
. the polymer weight by up to 9.9%, 8.3%, 5.1%, and 6.3% respectively for 90 .
days without any pre-treatment of ldPE. A slow and constantly
Replace with: … and an unclassified Bacillus sp. Pure cultures of the
. isolates reduced the polymer weight by up to 9.9%, 8.3%, 5.1%, and .
6.3% respectively after 90 days of growth without any pre-treatment of ldPE. In .
each case, a slow and constantly
Replaced
Page 13 Line 45: the strains at pH 11. The weight of the residual polymer reduced …..
Replace with: the participating strains at pH 11. The weight of the residual polymer was
. reduced …..
Replaced
Page 13 Lines 48
- 52: ….. isolated from the biofilm and the effectiveness of
. degradation by pure strains was investigated. A degree of 6.46% and
. 8.36% polymer weight loss was observed for the pure strains .
- correspondingly in the presence of IONPs and 9.62% and 11.32%,
- correspondingly in the presence of IONPs. Comparison of the
- Replace with: ….. isolated from the biofilm , both of which are classified as obligate.
. alkaliphilic Bacilliaceae. The effectiveness of degradation of PE by the pure strains
. was investigated. Corresponding polymer weight losses of 6.46% and 8.36%
. were observed for pure cultures of the isolates in the presence of IONPs , which
. increased to 9.62% and 11.32% in the presence of IONPs. Comparison of the
Replaced
Page 14 Lines1
- 5: degradation degree revealed highest efficiency for unformulated bacterial
. .consortium, followed by formulated consortium. Lowest efficiency was
. observed for individual strains.
. The identified alkaliphilic plastic degraders belong to the obligate
. alkaliphilic species of the family Bacilliaceae.
- Replace with: The same albeit enhanced trend was observed with both the
. unsupplemented and IONP-supplemented biofilm community, which even in the
. absence of the iron oxide nanoparticles was more effective than either of the
. strains isolated from the community.
Currently available information suggests that most dentified .
alkaliphilic plastic degraders are obligate alkaliphilic species of the family .
Bacilliaceae, a trend shared with the known thermophilic plastic degrading .
eubacteria.
Replaced
Page 14 Line 7
- 9: There is a lot of hope in the bioremediation of marine environments
. and salt niches like plastic polluted industrial wastewaters and salt
. marshes.
Replace with: There is growing evidence for the bioremediation of plastics in marine.
. and other natural saline environments such as salt marshes, as well as in
. salt-rich industrial wastewaters. Most of the characterised competent .
microorganisms have been found to be moderate or only slight .
extremophiles, with species of the genus Erythrobacter being .
predominant. Some examples of halophilic marine microorganisms .
able to degrade plastics are shown in Table 2. Significantly, the role of .
multi-species microbial biofilms in promoting plastic degradation in such .
environments has become increasingly recognised in recent years.
Replaced
Page 14 Line 10
- 15: The presence of different microbial ………….
………………PE (72.2%), followed by PP (18.0%), and PS
Replace with: A survey of different niches of seawater in the Western Mediterranean
. Sea [ ] consistently found that in each sampled area not only the highest
. number but also the highest density of bacteria were detected attached to
… plastic debris when compared with both sessile bacteria attached to .
other organic particles and free living bacteria. The plastic debris typically .
consisted of PE (72.2%), followed by PP (18.0%), and PS
Replaced
Page 14 Line16: ……. FTIR analysis. Dominant
Replace with: ……. FTIR analysis. By characterising the operational
. taxonomic units (OTUs) in the total DNA extracted from each of the
. samples, it was found that the dominant
Replaced
Page 14 Lines 20
- 25: 11.1%, respectively), while OTUs from organic particle ………….
…………………………. Large presence of Cyanobacteria
Replace with: 11.1% respectively) . In contrast, equivalent analyses confirmed that
the dominant microorganisms sourced from plastic debris were Cyanobacteria (40.8%,
mainly Pleurocapsa sp.) and Alphaproteobacteria (32.2%, mainly Roseobacter sp. and
Erythrobacter sp.), while the dominant microorganisms sourced from other organic
particles were Alphaproteobacteria (25.9%, mainly Erythrobacter sp.),
Gammaproteobacteria (25.0%, mainly Alteromonas sp.) and Cyanobacteria (17.9%,
mainly Synechococcus sp.). It was suggested that the relatively large recorded presence
of Cyanobacteria species on plastic debris was not only determined by their established
important role in biofilm formation [ * ], but also possibly by some activity towards plastic
debris: specifically identified strains belonged predominantly to two genera, Calothrix sp.
and Pleurocapsa which are known halophiles often isolated from marine environments.
Analysis of the OTUs sourced from plastic debris also confirmed that Erythrobacter
species were predominant (43%) amongst the detected hydrocarbonoclastic bacteria, and that two other moderate halophilic genera, Hyphomonas and Phorimidium, were present in significantly higher levels than in samples sourced from the other two environments.
Replaced
[ * ]. Additional new reference required to support this statement
Reference Rossi, F., & De Philippis, R., 2015 was included
Page 14 Line 36: Comparison on the ……. Replace with: Comparison of the …..
Replaced
Page 14 Line39: composition {54]. The established salinity suggests active growth of
Replace with: composition [54]. It was suggested that the significant difference in
. salinity may be reflected in the active growth of
Replaced
Page 14 Lines 42
- 43: the V3-V4 region of the 16S rRNA gene study of the microbial
. composition revealed different presented genera. Erythrobacter
- Replace with: the V3-V4 region of the 16S rRNA genes was used to characterise
. the microbial composition of the biofilms. Erythrobacter
Replaced
Page 14 Line 49
- 50: removal of biofilms during the traced time intervals resulted in a
. more intense biodegradation of the polymer. The dominant genera in PS
- Replace with: removal of biofilms throughout the 60-day incubation period resulted . in a more intense biodegradation of the polymer. In comparison, the , dominant genera in PS
Replaced
Page 15 Line 4
-10: the basis of the iVikodak package analysis. Based on the metabolic
. pathways of different taxa this enzyme was affiliated with the genera.
. Pseudomonas, Arenimonas, and Acidovorax in industrial water samples
. and Erythrobacter, Maribacter, and Mycobacterium in sea water samples
. As the same enzyme is involved in phenylalanine metabolism and as the
. mechanism of PS degradation is poorly understood, this hypothesis
. should be further confirmed.
Replace with: the basis of iVikodak metagenomics analysis [ * ]. Based on the
. known metabolic pathways of different taxa , this enzyme was most
. likely to be affiliated with detected species of the genera Pseudomonas,
., Arenimonas, and Acidovorax in the industrial water samples , and
. with detected species of the genera Erythrobacter, Maribacter, and
. Mycobacterium in the sea water samples. As the same enzyme is
. known to be involved in phenylalanine metabolism [ ** ], but its
. relationship to the mechanism of PS degradation is poorly understood,
. this requires further clarification.
Replaced
[ * ] Additional new reference required for this methodology
A reference Nagpal et al., 2019 was included
[ ** ] Additional new reference required
A reference Schmitt et al., 2017 was included
Page 15 Lines12
- 13: …… industrial waters with salinity 18.6, 0.09-0.3, and 1.3
. g/L , correspondingly [55]. Investigation of microbial diversity in these
. consortia revealed a universal ……..
- Replace with: …… industrial waters with respective salinity of 18.6, 0.09-0.3, and 1.3
. g/L [55]. Investigation of microbial diversity in consortia isolated from these
. environments revealed a universal ……..
Replaced
Page 15 Line 18: and Acidovorax due to the presence of genes responsible ….
Replace with: and Acidovorax due to the detected presence of genes responsible ….
Replaced
Page 15 Line 23: Biofilm formation on PCL and PVA …………
Replace with: The involvement of sessile microbial communities in the
. biodegradation of other types of plastic has been
. recognised. Biofilm formation on PCL and PVA …………
Replaced
Page 16 Lines 5
- 16: Approximately71% of the Earth is covered ……
. ……………………… microbal community composition and function .
[25]. Attachment to surface and growth within biofilms is an important
Replace with: Approximately 70% of the Earth is covered by marine water, 90% of
. which maintains a constant temperature of ~5oC regardless of latitude [57]. As a
. result, a predominant part of the Earth’s surface provides a suitable environment
. for the development of psychrophiles. Often marine microorganisms are multiple
. extremophiles, such as psychro-halophiles, psychro-piezophiles, or psychro- .
alkaliphiles [57]. Most of the plastic degrading bacteria identified in cold .
environments belong to the genera Shewanella, Moritella, and Psychrobacter. .
Some examples of psychrophilic marine microorganisms able to degrade .
plastics are shown in Table 2. While it has been proposed that .
such microorganisms play a role in degrading the increasing presence of .
plastics in marine ecosystems there is currently insufficient knowledge to .
understand the influence of plastics on microbial life, function, and community .
structure in these environments [25]. However, it is recognised that attachment to .
surfaces and growth within the resultant biofilms is an important
Replaced
Page 17 Line 7
- 16 ……………….(no more than 16oC) that suggests a -
……………………PU-A after 12 months incubation.
Replace with: ……………….(no more than 16oC) , and is thus favourable for the
. growth of psychrophilic bacteria. Consequently, it was chosen as the
. environment to compare the biodegradability of uncrosslinked
. poly(ethylene-butylene-adipate) (PU-A) and slightly crosslinked poly-
‘ (ε-caprolactone) (PU-B), two poly(ester-urethane) plastics with
. different structural characteristics [19]. Samples of both plastics were
. exposed to the seawater in Gdansk harbour (about 10oC and pH ~8). After 12 months a loss of tensile strength, discolouration, and cracking
. were recorded with both materials, but more significant for
. uncrosslinked PU-A.
Replaced
Page 17 Line 23: ….. is performed with higher rate compared to those in the Replace with: ….. is performed at a higher rate than in the
Replaced
Page 17 Line 29: Investigation on temperature and hydrostatic pressure effects on the Replace with: Investigation of the effects of temperature and hydrostatic pressure on the
Replaced
Page 17 Line 30: …………..revealed that Shewanella and Moritella …..
Replace with: revealed that all the Shewanella and Moritella …..
Replaced
Page 18 Table 2. This Table needs to be reformatted. Where it is not possible to include
. whole words, they need to be hyphenated in a ‘sensible’ way, eg
Degrad- Biodegrad- or Bio- Refer- or Ref-
ation able. degradable ence erence
Reformatted
Page 19 Line 4
- 5: ………………….. and growth profiles in
hydrostatic pressures an active ………
Replace with: ………………….. and growth profiles under different hydrostatic pressures , an active ………
Replaced
Page 19 Line 8; the cited reference for this research should be Oliveira et al 2021, not
. Oberbeckmann et al 2018
Corrected with Sekiguchi et al., 2011.
Page 19 Line 8: reliable way ……. Replace with: promising way ……
Replaced
Page 19 Line 21
- 22: ……Despite the short evolution time,
they have adapted ……………………….
Replace with: ……Despite the relatively short evolution time,
a significant number of extremophilic microorganisms have adapted ….
Replaced
Page 19.Line 26: …..their enzymes (the rigidity of the molecule limits the ….
Replace with: ………their enzymes (resulting from the rigidity of some plastic
. polymers which limits the ……
The phrase was clarified
Page 19 Line 30: this , i , the very extreme ……..
Replace with: this : firstly, the very extreme …..
Replaced
Page 19: Line31: ……………………….. degrade plastic; ii, the
Replace with: ………………………..degrade plastic , and secondly the
Replaced
Page 19 33: not be supported by such hard degradable polymers. Probably, after a
Replace with: not be totally supported by such hard to degrade polymers. It is possible
. that, after a ……
Replaced
Page 19 Lines 35
- 36: …………….is quite
. promising due to their ………………..
- Replace with: …………….. is a
- promising option due to their ………
This phrase was removed
Page 19 Line37: …. be developing of metagenome technology that permits ….
Replace with: …. be developing relevant metagenome technologies that permit ….
Replaced
Page 19 Line 38
- 39: … novel enzymes. or such with improved properties directly in
. environmental metagenomes, as well as ………
- Replace with: ……..novel enzymes or variants of known enzymes with improved
. relevant properties, as well as ……..
Replaced
Page 19 Line41: bacteria and their properties determined the object of the current …..
Replace with: bacteria and their properties was the objective of the current ……..
Replaced
Reviewer 2 Report
In this work, the authors present a review on plastic degradation by extremophilic bacteria. This is a very hot topic because plastic is one of the most significant pollution problems of our time.
First of all, they should have entered the number of lines to make the reviewers' job easier.
- General features of plastic degradation
I detect thought the text many formatting problems that the authors have to fix
In the Enzymes participating in plastic degradation part, the authors should provide the enzymes capable of degrading plastics with a more precise organization, also with the aid of a table, and also give some more precise examples of enzymes (derived from bacteria) capable of attacking plastics
paragraph 1.4 confuses me a bit. in the previous paragraph we talked about enzymatic activities, in this one instead of a "standard" method for the detection of biological activities against plastic. I believe the authors should reconsider this paragraph
- Extreme environments and extremophiles
In the first part of this paragraph, the authors should provide broader bibliographic support, providing examples of the most abundant microorganisms in extreme environments and provide some more indication of which kingdoms are found in these harsh environments.
Minor issues:
Fig 2 caption: close the ) after the reference.
- Alkaliphilic degraders, I think this point is 2.1 or 4, but in this case, you have to fix the numeration
Fig 5 всневщ ???? and provide the reference of this figure
Author Response
In this work, the authors present a review on plastic degradation by extremophilic bacteria. This is a very hot topic because plastic is one of the most significant pollution problems of our time.
First of all, they should have entered the number of lines to make the reviewers' job easier.
- General features of plastic degradation
I detect thought the text many formatting problems that the authors have to fix
I have checked carefully for formatting problems and hope will not appear again.
In the Enzymes participating in plastic degradation part, the authors should provide the enzymes capable of degrading plastics with a more precise organization, also with the aid of a table, and also give some more precise examples of enzymes (derived from bacteria) capable of attacking plastics
According to the reviewers recommendation a table (Table 1) was included. Enzymes were arranged with a more precise organization with an indication of the enzymes classes and bacterial sources.
paragraph 1.4 confuses me a bit. in the previous paragraph we talked about enzymatic activities, in this one instead of a "standard" method for the detection of biological activities against plastic. I believe the authors should reconsider this paragraph
Paragraph 1.4 place was changed (1.3) and enzyme assays discussed in the light of one of the standard methods for evaluation of biodegradation in the next paragraph.
- Extreme environments and extremophiles
In the first part of this paragraph, the authors should provide broader bibliographic support, providing examples of the most abundant microorganisms in extreme environments and provide some more indication of which kingdoms are found in these harsh environments.
Examples of the most abundant genera from the kingdom Bacteria were provided for each type of extreme environments supported by bibliographic information.
Minor issues:
Fig 2 caption: close the ) after the reference.
Done
- Alkaliphilic degraders, I think this point is 2.1 or 4, but in this case, you have to fix the numeration
The numeration was corrected
Fig 5 всневщ ???? and provide the reference of this figure
Slavic letters were removed
Reviewer 3 Report
I have no major criticism to this paper. It looks like a nice review. Given the glass transition temperature of many plastic polymers, thermophilic enzymes have emerged as promising biocatalysists for polymer degradation. The authors bring in other extreme environmental factors that may also affect the polymer structure (pH, salinity, etc.)
I can just point at some suggested changes/amendments.
- The authors briefly refer to acidophiles and piezophiles in this review and the lack of reports for plastic degradation in these environments and therefore, they are not covered. Given that both pressure and pH (and oxidative stress in some extreme acidic environments) may also contribute to the polymer estability, do they authors foresee any relevant findings coming from these type of environments in the near future?
- Maybe the authors want to make a reference to microplastics when they talk about biodeterioration.
- When talking about groups of enzymes, I would probably firstly refer to enzyme classes (hydrolases and oxidoreductases) and then mention the groups. This is because both esterases, proteases and cutinases are all hydrolases. Moreover, cutinase is a special type of esterase.
- Eukaryote is probably the right form (not eucaryote).
- Maybe you want to refer to "facultative thermophiles" as "thermotolerant"?
- Other relevant thermophilic environments include the deep terrestrial subsurface, or even dry arid soils can host thermophiles. As for low temperature environments, I would probably mention sea ice or deep sea waters as very relevant for this subject.
- "subsp" is normally not in italics (check both Table 1 and main text).
- If you include the phylum/class of all others, then both Arenimonas and Pseudomonas belong to Gammaprotebacteria.
- Phenylacetaldehyde dehydrogenase is EC 1.2.1.39
- Black Sea, both words are capitalized.
- Consider to use the full name the first time you name UNEP.
- Table 2. The column "effectiveness of degradation" should be completed. If it is not reported, consider to write "not reported".
- The authors should consider citing the work by Sulaiman et al., 2012, where the report an polyester hydrolase from a metagenome obtained from compost, which is also slightly thermoestable (10.1128/AEM.06725-11).
Author Response
I have no major criticism to this paper. It looks like a nice review. Given the glass transition temperature of many plastic polymers, thermophilic enzymes have emerged as promising biocatalysists for polymer degradation. The authors bring in other extreme environmental factors that may also affect the polymer structure (pH, salinity, etc.)
I can just point at some suggested changes/amendments.
- The authors briefly refer to acidophiles and piezophiles in this review and the lack of reports for plastic degradation in these environments and therefore, they are not covered. Given that both pressure and pH (and oxidative stress in some extreme acidic environments) may also contribute to the polymer estability, do they authors foresee any relevant findings coming from these type of environments in the near future?
The next sentences were included:
“…are not available although the decreased plastic strength in acidic environment suggests a good prospect for faster waste degradation. In the same time acidic pH shorten life of some plastic products used in bleaching processes.”
and
“In the same time a decrease in plastic strength by increased pressure at depth water should be consider.”
- Maybe the authors want to make a reference to microplastics when they talk about biodeterioration.
Microplastics with their peculiarities in the mechanism of degradation are not an object of the current review.
- When talking about groups of enzymes, I would probably firstly refer to enzyme classes (hydrolases and oxidoreductases) and then mention the groups. This is because both esterases, proteases and cutinases are all hydrolases. Moreover, cutinase is a special type of esterase.
Enzymes were arranged with an indication of the enzymes classes and then their groups in Table 1.
- Eukaryote is probably the right form (not eucaryote).
Done
- Maybe you want to refer to "facultative thermophiles" as "thermotolerant"?
The next sentences were included:
“Facultative thermophiles thrive at temperatures 41-50 °C, while thermotolerant microorganisms are mesophilic microorganisms that can tolerate temperature higher than 41°C however grow optimally at lower temperature.”
- Other relevant thermophilic environments include the deep terrestrial subsurface, or even dry arid soils can host thermophiles. As for low temperature environments, I would probably mention sea ice or deep sea waters as very relevant for this subject.
As an information concerning plastic degradation in the deep terrestrial subsurface, dry arid soils and sea ice was not found, they were not included in the review. The next phrase was included in the sentence:
“Cold environments comprise fresh and marine waters including deep sea water,…”
- "subsp" is normally not in italics (check both Table 1 and main text).
Corrected
- If you include the phylum/class of all others, then both Arenimonasand Pseudomonas belong to Gammaprotebacteria.
Included
- Phenylacetaldehyde dehydrogenase is EC 1.2.1.39
Added
- Black Sea, both words are capitalized.
Corrected
- Consider to use the full name the first time you name UNEP.
Full name “United Nations Environment Programme” was used in the first mention.
- Table 2. The column "effectiveness of degradation" should be completed. If it is not reported, consider to write "not reported".
Table 2 was completed.
- The authors should consider citing the work by Sulaiman et al., 2012, where the report an polyester hydrolase from a metagenome obtained from compost, which is also slightly thermoestable (10.1128/AEM.06725-11).
Cutinase work of Sulaiman et al., 2012 was included.
Reviewer 4 Report
I really liked this well-written and comprehensive review on plastic degradation by extremophilic bacteria. This is a critical subject for environmental studies now days. Unfortunately, the data on this matter is scarce, the processes are understudied and very far from real understanding. Yet, the input and impact of plastics in the environment are enormous.
The Authors did a great job collecting all available data on plastics degradation in extreme environments, such as high and low temperatures, high alkalinities and high salt. It could be very helpful and might promote research in this field.
I have some minor comments mostly regarding English language.
Paragraph 1.2 “relative deal of crystalline and amorphous…” Use “share” instead of deal
“size and shape of substrates” use “form” instead of shape
“plastics are divided into two main groups” use colon (:) instead of comer.
Use comer before “which”
“are further degraded by secondary degraders into their cells” Remove “into their cells”
“oligomers” after dash – small letter.
“approach for resolving of plastic disposal problems” remove “of”
“microorganisms; gene expression…” Use “or” instead of semicolon
Paragraph 1.3
“the resistance of plastics against microbial attack” Use resistance to microbial attack
“ but to the attack to the chemical additives in their molecules” Is it “attack OF chemical additives ON their molecules”? Not clear sentence.
Paragraph 1.4
“5. Registration of products” It is better to use “detection” instead of “registration”.
“showed different success” It’s better to use “variable” instead of “different”.
“formed biofilms which was related with degradation of PE, biofilms by….”
Use “…that were related to degradation of PE.” A dot here. Then start a new sentence
and use either “biofilms formed BY” or “biofilms of Ps. Xxxx”. Start again a new sentence on B. subtilis and B. cereus.
Part 2.
“Growth of hyperthermophiles is best at above 80C [31]. Is it right reference? Check ,please!
The same about reference [32].
“osmotic pressure balance reveived byorganic compatable solutes” Did you mean “achieved”?
Again, check the ref. [32].
Table 1 and Table 2. First column should be called “Plastic degradation type”. Degradation type refers to a chemical reaction.
“Sequence differences” Did you mean aminoacids sequence differences? Should be clarified.
“two the best of our knowledge” is used two times. Remove, please.
Wrong numbering of the chapters. “Alkaliphilic degraders “is numbered 1. Yet, the previous chapter on Extremophiles was numbered 2. Make the numbering uniformed throughout the paper, please!
“The addition of IONPs facilitated biofilm formation.” Is it any explanation to this?
4.Halophilic degraders.
“Two other moderate halophilic genera…… were 10-100 more found….”
Use “Two other moderately hallophiliuc genera were 10-100 more abundant”
“ in comparison with other two communities” Its better to clarify what communities than to send a reader to go looking for that in the paper.
“iVikodak package analysis” requires a link in brackets.
Like other extremophiles halophilic producers of plastic degrading enzymes are slight and moderate extremophiles” requires a comer after “other extremophiles”
The above statement sounds like a conclusion. In this respect, next sentence looks out of place. Not logical.
Conclusions
Use “human” instead of “man life”.
Author Response
I really liked this well-written and comprehensive review on plastic degradation by extremophilic bacteria. This is a critical subject for environmental studies now days. Unfortunately, the data on this matter is scarce, the processes are understudied and very far from real understanding. Yet, the input and impact of plastics in the environment are enormous.
The Authors did a great job collecting all available data on plastics degradation in extreme environments, such as high and low temperatures, high alkalinities and high salt. It could be very helpful and might promote research in this field.
I have some minor comments mostly regarding English language.
Paragraph 1.2 “relative deal of crystalline and amorphous…” Use “share” instead of deal
Done
“size and shape of substrates” use “form” instead of shape
Done
“plastics are divided into two main groups” use colon (:) instead of comer.
Done
Use comer before “which”
Done
“are further degraded by secondary degraders into their cells” Remove “into their cells”
Removed
“oligomers” after dash – small letter.
Done
“approach for resolving of plastic disposal problems” remove “of”
Done
“microorganisms; gene expression…” Use “or” instead of semicolon
Done
Paragraph 1.3
“the resistance of plastics against microbial attack” Use resistance to microbial attack
Done
“ but to the attack to the chemical additives in their molecules” Is it “attack OF chemical additives ON their molecules”? Not clear sentence.
The phrase was changed next way:
“…to the attack of the chemical additives.”
Paragraph 1.4
“5. Registration of products” It is better to use “detection” instead of “registration”.
Changed
“showed different success” It’s better to use “variable” instead of “different”.
Changed
“formed biofilms which was related with degradation of PE, biofilms by….”
Use “…that were related to degradation of PE.” A dot here. Then start a new sentence
and use either “biofilms formed BY” or “biofilms of Ps. Xxxx”. Start again a new sentence on B. subtilis and B. cereus.
Done
Part 2.
“Growth of hyperthermophiles is best at above 80C [31]. Is it right reference? Check ,please!
Corrected
The same about reference [32].
Corrected
“osmotic pressure balance reveived byorganic compatable solutes” Did you mean “achieved”?
Changed
Again, check the ref. [32].
Table 1 and Table 2. First column should be called “Plastic degradation type”. Degradation type refers to a chemical reaction.
Corrected
“Sequence differences” Did you mean aminoacids sequence differences? Should be clarified.
Aminoacid was added
“two the best of our knowledge” is used two times. Remove, please.
Removed
Wrong numbering of the chapters. “Alkaliphilic degraders “is numbered 1. Yet, the previous chapter on Extremophiles was numbered 2. Make the numbering uniformed throughout the paper, please!
Numbering was corrected.
“The addition of IONPs facilitated biofilm formation.” Is it any explanation to this?
The authors explanation was included:
The effect of IONPs was attributed to the properties of the nanoparticles such as magnetism and electrostatic charge leading to alter bacterial motion through signal transduction as well as the formation of cofactors produced in the medium. As a result higher hydrophobicity of the consortium with IONPs and higher adhesion to the plastic surface was demonstrated.
4.Halophilic degraders.
“Two other moderate halophilic genera…… were 10-100 more found….”
Use “Two other moderately hallophiliuc genera were 10-100 more abundant”
Changed
“ in comparison with other two communities” Its better to clarify what communities than to send a reader to go looking for that in the paper.
Clarified
“iVikodak package analysis” requires a link in brackets.
A reference was included.
Like other extremophiles halophilic producers of plastic degrading enzymes are slight and moderate extremophiles” requires a comer after “other extremophiles”
The above statement sounds like a conclusion. In this respect, next sentence looks out of place. Not logical.
This paragraph was changed next way:
The moderate halophilic representatives of the genus Erythrobacter are often identified as an actively growing in plastic. Like other extremophiles, halophilic producers of plastic degrading enzymes are slight and moderate extremophiles.
Conclusions
Use “human” instead of “man life”.
Done
Round 2
Reviewer 1 Report
The following need attention:
a. Page 8: The concluding paragraph of section 1.3.2. (Enzymes participating in ... etc) should include the addition previously suggested, ie:
It is known that extremophilic microorganisms are competent producers of a range of potentially relevant hydrolytic enzymes[Kour et al 2019].
The reference 'Kour et al 2019' will now become [42], and all current references [42] et seq will need to be renumbered accordingly as [43] et seq both in text and in the Bibliography.
The full details of this reference are:
'Extremophiles for hydrolytic enzyme production: biodiversity and potential biotechnological applications'
D.Kour, K.L.Rana, T.Kour, B.Singh, V.S.Chauhan, A.Kumar, A.A.Rastegan, N.Yadav, A.J.Nadav, and V.K.Gupta.
pp321 -372 in Bioprocessing for Biomolecules Production
Edited by G.Molina, V.K.Gupta, B.N.Singh, and N.Gathergood.
John Wiley & Sons, November 2019
doi:10.1002/9781119434436.ch16
b. Page 12, Table 2: i. Mw replace with MW (one example of this remains in text). ii. Polycaprolacto ne replace with Polycapro lactone
c. Page 19, Table 3: Plastic degradatio n type replace with Plastic degradation type. Ideally, should also replace Biodegr arable with Biodegrad able, but this will possibly require reformatting the Table
Author Response
- Page 8: The concluding paragraph of section 1.3.2. (Enzymes participating in ... etc) should include the addition previously suggested, ie:
It is known that extremophilic microorganisms are competent producers of a range of potentially relevant hydrolytic enzymes[Kour et al 2019].
The reference 'Kour et al 2019' will now become [42], and all current references [42] et seq will need to be renumbered accordingly as [43] et seq both in text and in the Bibliography.
The full details of this reference are:
'Extremophiles for hydrolytic enzyme production: biodiversity and potential biotechnological applications'
D.Kour, K.L.Rana, T.Kour, B.Singh, V.S.Chauhan, A.Kumar, A.A.Rastegan, N.Yadav, A.J.Nadav, and V.K.Gupta.
pp321 -372 in Bioprocessing for Biomolecules Production
Edited by G.Molina, V.K.Gupta, B.N.Singh, and N.Gathergood.
John Wiley & Sons, November 2019
doi:10.1002/9781119434436.ch16
The reference 42 was included and the numbers of the next references were changed.
- Page 12, Table 2: i. Mw replace with MW (one example of this remains in
text).
Mw was replaced by MW
- Polycaprolacto ne replace with Polycapro lactone
The used Page setup for Table 2 and 3 in the variant I sent was Landscape and the word Polycaprolactone is on one row. I cannot understand why the word is separate. That’s why I am sending now also pdf format.
- Page 19, Table 3: Plastic degradatio n type replace with Plastic degradation type. Ideally, should also replace Biodegr arable with Biodegrad able, but this will possibly require reformatting the Table
The used Page setup for Table 2 and 3 in the variant I sent was Landscape and the word Polycaprolactone is on one row. I cannot understand why the word is separate. That’s why I am sending now also pdf format.
Reviewer 2 Report
The authors adequately revised the paper. it is my opinion that it can be accepted in the current version
Author Response
The authors adequately revised the paper. it is my opinion that it can be accepted in the current version.
As the reviewer has no other comments except paper acceptance, I support his suggestion.
