Author:
Xiao Han,Liu Xiufang,Feng Yunzi,Zheng Lin,Zhao Mouming,Huang Mingtao
Abstract
Abstract
Background
The production and processing of animal-based products generates many collagen-rich by-products, which have received attention both for exploitation to increase their added value and to reduce their negative environmental impact. The collagen-rich by-products can be hydrolyzed by collagenases for further utilization. Therefore, collagenases are of benefit for efficient collagen materials processing. An alternative and safe way to produce secreted collagenases is needed.
Results
Two collagenases from Hathewaya histolytica, ColG and ColH, were successfully secreted by the yeast Saccharomyces cerevisiae. Compared with the native signal peptide of collagenase, the α-factor leader is more efficient in guiding collagenase secretion. Collagenase secretion was significantly increased in YPD medium by supplementing with calcium and zinc ions. Recombinant collagenase titers reached 68 U/mL and 55 U/mL for ColG and ColH, respectively. Collagenase expression imposed metabolic perturbations on yeast cells; substrate consumption, metabolites production and intracellular cofactor levels changed in engineered strains. Both recombinant collagenases from yeast could hydrolyze soluble and insoluble collagen materials. Recombinant ColG and ColH showed a synergistic effect on efficient collagen digestion.
Conclusions
Sufficient calcium and zinc ions are essential for active collagenase production by yeast. Collagenase secretion was increased by optimization of expression cassettes. Collagenase expression imposed metabolic burden and cofactor perturbations on yeast cells, which could be improved through metabolic engineering. Our work provides a useful way to produce collagenases for collagen resource utilization.
Graphical Abstract
Funder
Key Research and Development Program of Guangdong Province, China
National Natural Science Foundation of China
Science and Technology Program of Guangzhou, China
Publisher
Springer Science and Business Media LLC
Subject
Management, Monitoring, Policy and Law,Energy (miscellaneous),Applied Microbiology and Biotechnology,Renewable Energy, Sustainability and the Environment,Biotechnology
Cited by
6 articles.
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