Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
3.7
3.7
Journals
Fermentation
Special Issues
Biotechnological Potential of Escherichia coli and Pichia pastoris: Expression...
share announcement

Biotechnological Potential of Escherichia coli and Pichia pastoris: Expression of Recombinant Proteins and Synthesis of Interesting Molecules

A special issue of Fermentation (ISSN 2311-5637). This special issue belongs to the section "Microbial Metabolism, Physiology & Genetics".

Deadline for manuscript submissions: closed (31 January 2023) | Viewed by 5703

Special Issue Editor

Dr. Xiaoqing Hu

E-Mail Website
Guest Editor
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China
Interests: metabolic engineering; fermentation; Pichia pastoris; chassis design; Escherichia coli; biosynthesis; lipopolysaccharide modification; metabolic regulation; food microbiology

Special Issue Information

Dear Colleagues,

Industrial strains of bacteria and yeast, especially Escherichia coli and Pichia pastoris, have utilized renewable feedstock for the expression of recombinant proteins and biosynthesis of a broad range of interesting chemicals. Today, along with the development of biotechnologies such as genome editing and multi-omics, a variety of strategies in molecular biology, such as chassis redesign, genome minimization, cell membrane engineering, promoter engineering, directed evolution of protein, RNA interference, and the redesign of biosynthesis pathways, are being employed to improve the performance of industrial strains.  

This Special Issue focuses on these optimization strategies on different scales (such as gene, protein, RNA, cofactor, pathway, and host cell) to improve recombinant proteins and the synthesis of interesting molecules in E. coli and P. Pastoris. Additionally, the fermentation optimization of these modified strains at the bioreactor level is also welcome. For this reason, this issue welcomes highly interdisciplinary studies from disparate research fields, and original research articles and reviews are accepted.

Dr. Xiaoqing Hu
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Fermentation is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • genome minimization
  • chassis design
  • synthetic biology
  • membrane engineering
  • Pichia pastoris
  • Escherichia coli
  • heterologous expression
  • biosynthesis
  • fermentation optimization
  • metabolic regulation

Published Papers (3 papers)

Download All Papers
Order results
Result details
Show export options expand_more Show export options expand_less
Select all
Export citation of selected articles as:

Research

14 pages, 4514 KiB  
Article
Enhancing the Activity of a Self-Inducible Promoter in Escherichia coli through Saturation Mutation and High-Throughput Screening
by Jinyang Li, Sheng Tong, Farrukh Raza Amin, Habiba Khalid, Kai Chen, Xiaoguang Zhao, Jinling Cai and Demao Li
Fermentation 2023, 9(5), 468; https://doi.org/10.3390/fermentation9050468 - 13 May 2023
Cited by 1 | Viewed by 1431
Abstract
The use of self-inducible promoters is a promising strategy to address metabolic imbalances caused by overexpression. However, the low activity of natural self-inducible promoters hinders their widespread application. To overcome this limitation, we selected the fic promoter as a model promoter to create [...] Read more.
The use of self-inducible promoters is a promising strategy to address metabolic imbalances caused by overexpression. However, the low activity of natural self-inducible promoters hinders their widespread application. To overcome this limitation, we selected the fic promoter as a model promoter to create an enhanced self-inducible promoter library using saturation mutations and high-throughput screening. Sequence analysis revealed that these promoters share certain characteristics, including semi-conservation in the −35 hexamer, highly conserved cytosine in the −17 motif (compared to −13 for other promoters), and moderate A+T content between positions −33 and −18 in the spacer region. Additionally, the discriminator region of these promotors features high A+T content in the first five bases. We identified PficI-17, PficII-33, and PficIII-14 promoters as the optional promoters in the −35 hexamer, spacer region, and discriminator mutation libraries, respectively. These promotors were used as representatives to measure the specific fluorescence and OD600 nm dynamics in different media and to confirm their effect on the expression of different proteins, including egfp (enhanced green fluorescence protein) and rfp (red fluorescence protein). Overall, our findings provide valuable guidance for modifying promoters and developing a promoter library suitable for regulating target genes. Full article
(This article belongs to the Special Issue Biotechnological Potential of Escherichia coli and Pichia pastoris: Expression of Recombinant Proteins and Synthesis of Interesting Molecules)
►▼ Show Figures

Figure 1

13 pages, 1515 KiB  
Article
Enhancing Antimicrobial Peptide Productivity in Pichia pastoris (Muts Strain) by Improving the Fermentation Process Based on Increasing the Volumetric Methanol Consumption Rate
by Chatchol Kongsinkaew, Supenya Chittapun, Chanitchote Piyapittayanun, Viroj Boonyaratanakornkit, Sarintip Sooksai, Kittisak Ajariyakhajorn, Soisuda Pornpukdeewattana, Warawut Krusong, Tunyaboon Laemthong and Theppanya Charoenrat
Fermentation 2023, 9(3), 277; https://doi.org/10.3390/fermentation9030277 - 12 Mar 2023
Cited by 4 | Viewed by 2443
Abstract
The instability of the protein expression in Pichia pastoris strains has been an issue for various peptide productions. Some modifications to the traditional fermentation process could potentially solve the problem. Here, we consider a four-stage fermentation process to express the CAP2 (cell-penetrating antimicrobial [...] Read more.
The instability of the protein expression in Pichia pastoris strains has been an issue for various peptide productions. Some modifications to the traditional fermentation process could potentially solve the problem. Here, we consider a four-stage fermentation process to express the CAP2 (cell-penetrating antimicrobial peptide 2) candidate in P. pastoris KM71H, a slow methanol utilization strain. During the fermentation process, CAP2 productivity is limited (6.15 ± 0.21 mg/L·h) by the low overall methanol consumption (approximately 645 g), which is mainly the result of the slow methanol utilization of the P. pastoris KM71H. To overcome this limitation, we increased the cell concentration two-fold prior to the induction stage. A fed-batch process with exponential and dissolved oxygen tension (DOT) stat feeding strategies was deployed to control the glycerol feed, resulting in an increase in cell concentration and enhancement of the volumetric methanol consumption rate. The improved fermentation process increased the overall methanol consumption (approximately 1070 g) and the CAP2 productivity (13.59 ± 0.24 mg/L·h) by 1.66 and 2.21 times, respectively. In addition, the CAP3 (cell-penetrating antimicrobial peptide 3) candidate could also be produced using this improved fermentation process at a high yield of 3.96 ± 0.02 g/L without any further optimization. Note that there was no oxygen limitation during the improved fermentation process operating at high cell density. This could be due to the controlled substrate addition via the DOT stat system. Full article
(This article belongs to the Special Issue Biotechnological Potential of Escherichia coli and Pichia pastoris: Expression of Recombinant Proteins and Synthesis of Interesting Molecules)
►▼ Show Figures

Figure 1

16 pages, 3294 KiB  
Article
Improving the Synthesis Efficiency of Amino Acids Such as L-Lysine by Assembling Artificial Cellulosome Elements Dockerin Protein In Vivo
by Nan Li, Le Xue, Zirui Wang, Peng Du, Piwu Li, Jing Su, Jing Xiao, Min Wang, Junqing Wang and Ruiming Wang
Fermentation 2022, 8(11), 578; https://doi.org/10.3390/fermentation8110578 - 25 Oct 2022
Cited by 2 | Viewed by 1198
Abstract
Cellulosome is a highly efficient multi-enzyme self-assembly system and is found on the extracellular surface or in the free environment of microorganisms. However, with a lack of Ca2+ in vivo, cellulosome assembly is challenging. In this study, a novel design method was [...] Read more.
Cellulosome is a highly efficient multi-enzyme self-assembly system and is found on the extracellular surface or in the free environment of microorganisms. However, with a lack of Ca2+ in vivo, cellulosome assembly is challenging. In this study, a novel design method was used to directionally modify the Ca2+-binding site, and four double-site dockerin A (DocA) mutants were obtained. At a Ca2+ concentration between 1.00 × 10−7 and 1.00 × 10−4 M, the mutant DocA-D3 had the strongest binding capacity to cohesion (Coh), which was 8.01 times that of DocA. The fluorescence signal intensity of the fusion proteins assembled using mutants was up to 1.26 × 107 in Escherichia coli, which indicated that these mutants could interact with Coh in vivo. The molecular dynamics simulation results showed that DocA-D3 could maintain a stable angle structure without Ca2+, and when applied to L-lysine fermentation, the yield was increased by 24.1%; when applied to β-alanine fermentation, the product accumulation was increased by 2.13–2.63 times. These findings lay the foundation for assembly design in cells. Full article
(This article belongs to the Special Issue Biotechnological Potential of Escherichia coli and Pichia pastoris: Expression of Recombinant Proteins and Synthesis of Interesting Molecules)
►▼ Show Figures

Figure 1

Show export options expand_more Show export options expand_less
Select all
Export citation of selected articles as:
 
Displaying articles 1-3
Back to TopTop