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Homologous High-Level Lipase and Single-Cell Protein Production with Engineered Yarrowia lipolytica via Scale-Up Fermentation for Industrial Applications

Fermentation 2023, 9(3), 268; https://doi.org/10.3390/fermentation9030268

by Dujie Pan^1,†, Shuhan Dai^1,†, Liangcheng Jiao¹, Qinghua Zhou¹, Genhan Zha¹, Jinyong Yan¹, Bingnan Han²

, Yunjun Yan¹

and Li Xu^1,*

Reviewer 1: Anonymous

Reviewer 2: Anonymous

Fermentation 2023, 9(3), 268; https://doi.org/10.3390/fermentation9030268

Submission received: 23 February 2023 / Revised: 5 March 2023 / Accepted: 7 March 2023 / Published: 9 March 2023

(This article belongs to the Special Issue Yarrowia lipolytica: A Beneficial Yeast as a Biofactory for Biotechnological Applications)

Round 1

Reviewer 1 Report

The topic of the paper „Homologous high-level lipase and single-cell protein production from engineering Yarrowia lipolytica via scale-up fermentation for industrial applications” is very interesting for readers, Yarrowia lipolytica being a promising source of novel feed additives.

The aim of this study was to produce extracellular lipases and single-cell proteins at high levels simultaneously through fed-batch fermentation of engineered Yarrowia lipolytica.

The results obtained showed a balance between lipase secretion and cell growth of the Yarrowia lipolytica recombinant strain, indicating that an efficient fermentation strategy could promote further scale-up for industrial single-cell proteins production from homologous engineered Yarrowia lipolytica.

The introduction provides sufficient background and includes relevant references.

The manuscript is well written, and the text is easy to read.

The design research is well described.

The results are consistent and clearly presented.

The reference list is variously and relatively recently.

Author Response

Thank you for your valuable suggestion! This is the best encouragement. We greatly appreciate your time and your meticulous review of our manuscript. Your positive comments are the best encouragement. We look forward to hearing from you.

Best regards.

Sincerely,

Li Xu

Key Laboratory of Molecular Biophysics of the Ministry of Education

College of Life Science and Technology

Huazhong University of Science and Technology

Postal address: Luoyu Road 1037, Wuhan, China

Wuhan 430074, P. R. China

Telephone number: +86-27-8779-2214

Fax number: +86-27-8779-2214

E-mail address: xuli@hust.edu.cn

Author Response File: Author Response.docx

Reviewer 2 Report

The work is within the scope of the journal presents excellent research that can be published after the slightest revision is done. You need to improve the quality of the Figures, as in many it is not possible to see the values on the axes. Compare your results with others in the literature, including SDS-PAGE and different sources of lipases. A simple search for lipase in Science Direct easily finds 150,000 articles, of which 30,000 in the last 5 years. I know your source is different but you need to justify this better in the introduction (more than 20 articles) and defend its potential in relation to existing ones in the results and discussion

Author Response

Thank you for your valuable suggestion and encouragement! The major revisions are outlined in detail below.

The quality of Figures has been improved by enhancing the DPI values and enlarged the fonts of the values on the axes, which were inserted in the text of the revised version of this manuscript.
According to the suggestion, our results is further compared with others in the cited literature, including the results of SDS-PAGE listed following “…… In Figures 5–6, there was a main band with a molecular mass of approximately 38 kDa in the supernatant from fed-batch fermentation, which is the molecular mass of Y. lipolytica lipase (YLL) Lip2 [29] ……” in the part of RESULTS.
As you mentioned, lipases have attracted increasing attentions and become one research hot spots because their unique properties and versatile application in the fields of food, flavor, pharmaceutical, leather, textile, cosmetic, paper industries, energy and environment (Ali et al., 2023; Akram et al., 2023; Cao et al., 2022; Groenewald et al., 2014; Liu et al., 2015; Madzak et al., 2018; Mahfoudhi et al., 2022; Patel et al., 2019; Reetz MT 2002, 2004, 2012, and 2013; Turner et al., 2009). There are so many reports about lipases including source screening, overexpression, and modification by engineering aimed to obtain plenty of green biocatalyst. However, the utilization of lipases as helper of oil and fat digestion in the field of feed has been relatively ignored. It is promising strategy to produce value-added products using waste substrates from an industrial point of view. For feed purpose, the recombinant auxotrophic Y. lipolytica strains YLY1, YLY2, YLY3, YLY4, and YLY5 were engineered for simultaneous production of lipase and SCP on cost-effective media by synthetic biological strategy. Fish oral feeding assessment further demonstrated that this engineered yeast YLY5 were an excellent feed additives (Yan et al., 2018). Therefore, scaling up processes for the simultaneous and efficient production of lipase and cell mass economically is significant aimed at potential feed industrial applications. In this context, the high-density fermentation of best engineering Y. lipolytica strain were further investigated from agro-industrial wastes for feed in batch fermentation.
Thanks for the kind reminder. More recently references have been cited and their numbers have regulated accordingly. The related reports about lipase are as Reference 3, 8, 13, 14, 15, 16, 17, 29, 36 are cited in the revised version. There are 26 articles to be cited in the part of INTRODUCTION.

We greatly appreciate for your time and your meticulous review of our manuscript. Your positive comments are the best encouragement. We look forward to hearing from you.