Construction of efficient xylose utilizingPichia pastoris for industrial enzymeproduction

Microbial Cell Factories

Research

Pengfei Li¹  Taicheng Zhu¹  Yin Li¹  Zao Chen²  Hongbing Sun³ 

[1] CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China;CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China;Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, 361005, Xiamen, China;National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China;

关键词: Xylose;    Xylose isomerase;    Pichia pastoris;    Evolutionary engineering;    Industrial enzymes;   

DOI  :  10.1186/s12934-015-0206-8

 received in 2014-09-22, accepted in 2015-02-06,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundCellulosic biomass especially agricultural/wood residues can beutilized as feedstock to cost-effectively produce fuels, chemicals and bulkindustrial enzymes, which demands xylose utilization from microbial cellfactories. While previous works have made significant progress in improvingmicrobial conversion of xylose into fuels and chemicals, no study has reportedthe engineering of efficient xylose utilizing protein expression systems for thepurpose of producing industrial enzymes.ResultsIn this work, using Pichiapastoris as an example, we demonstrated the successfulengineering of xylose metabolizing ability into of protein expression systems. Aheterologous XI (xylose isomerase) pathway was introduced into P. pastoris GS115 by overexpressing the Orpinomyces spp. XI or/and the endogenous XK(xylulokinase) gene, and evolutionary engineering strategies were also applied.Results showed that the XI pathway could be functionally expressed in P. pastoris. After 50 generation of sequentialbatch cultivation, a set of domesticated recombinant P.pastoris strains with different performance metrics on xylosewere obtained. One evolved strain showed the highest xylose assimilationability, whose cell yield on xylose can even be comparable to that on glucose orglycerol. This strain also showed significantly increased β-mannanase productionwhen cultured on xylose medium. Furthermore, transcription analysis of xylosepathway genes suggested that overexpression of XI and XK might be the keyfactors affecting effective xylose assimilation.ConclusionsTo our best knowledge, this study is the first work demonstratingthe construction of efficient xylose utilizing P.pastoris strains, thus providing a basis for using cellulosicbiomass for bulk industrial enzyme production.

【 授权许可】

Unknown   
© Li et al.; licensee BioMed Central. 2015. This is an Open Access article distributed under the terms of theCreative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproductionin any medium, provided the original work is properly credited. The CreativeCommons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unlessotherwise stated.