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Engineering Flocculation for Improved Tolerance and Production of d-Lactic Acid in Pichia pastoris
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Document Title
Engineering Flocculation for Improved Tolerance and Production of d-Lactic Acid in Pichia pastoris
Author
Sae-Tang K., Bumrungtham P., Mhuantong W., Champreda V., Tanapongpipat S., Zhao X.-Q., Liu C.-G., Runguphan W.
Affiliations
National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathum Thani, 12120, Thailand; State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China
Type
Article
Source Title
Journal of Fungi
ISSN
2309608X
Year
2023
Volume
9
Issue
4
Page
-
Open Access
All Open Access, Gold, Green
Publisher
MDPI
DOI
10.3390/jof9040409
Format
Abstract
d-lactic acid, a chiral organic acid, can enhance the thermal stability of polylactic acid plastics. Microorganisms such as the yeast Pichia pastoris, which lack the natural ability to produce or accumulate high amounts of d-lactic acid, have been metabolically engineered to produce it in high titers. However, tolerance to d-lactic acid remains a challenge. In this study, we demonstrate that cell flocculation improves tolerance to d-lactic acid and increases d-lactic acid production in Pichia pastoris. By incorporating a flocculation gene from Saccharomyces cerevisiae (ScFLO1) into P. pastoris KM71, we created a strain (KM71-ScFlo1) that demonstrated up to a 1.6-fold improvement in specific growth rate at high d-lactic acid concentrations. Furthermore, integrating a d-lactate dehydrogenase gene from Leuconostoc pseudomesenteroides (LpDLDH) into KM71-ScFlo1 resulted in an engineered strain (KM71-ScFlo1-LpDLDH) that could produce d-lactic acid at a titer of 5.12 ± 0.35 g/L in 48 h, a 2.6-fold improvement over the control strain lacking ScFLO1 expression. Transcriptomics analysis of this strain provided insights into the mechanism of increased tolerance to d-lactic acid, including the upregulations of genes involved in lactate transport and iron metabolism. Overall, our work represents an advancement in the efficient microbial production of d-lactic acid by manipulating yeast flocculation. © 2023 by the authors.
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
Ministry of Higher Education, Science, Research and Innovation, Thailand
Publication Source
WOS