Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation
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...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-08-01
|
| Series: | Biotechnology for Biofuels and Bioproducts |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-022-02186-y |
| _version_ | 1797752817262264320 |
|---|---|
| author | Han Xiao Xiufang Liu Yunzi Feng Lin Zheng Mouming Zhao Mingtao Huang |
| author_facet | Han Xiao Xiufang Liu Yunzi Feng Lin Zheng Mouming Zhao Mingtao Huang |
| author_sort | Han Xiao |
| collection | DOAJ |
| description | 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 |
| first_indexed | 2024-03-12T17:08:56Z |
| format | Article |
| id | doaj.art-1c7bfe926f0245eba1c615e299c51294 |
| institution | Directory Open Access Journal |
| issn | 2731-3654 |
| language | English |
| last_indexed | 2024-03-12T17:08:56Z |
| publishDate | 2022-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj.art-1c7bfe926f0245eba1c615e299c512942023-08-06T11:09:32ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542022-08-0115111310.1186/s13068-022-02186-ySecretion of collagenases by Saccharomyces cerevisiae for collagen degradationHan Xiao0Xiufang Liu1Yunzi Feng2Lin Zheng3Mouming Zhao4Mingtao Huang5School of Food Science and Engineering, South China University of TechnologySchool of Food Science and Engineering, South China University of TechnologySchool of Food Science and Engineering, South China University of TechnologySchool of Food Science and Engineering, South China University of TechnologySchool of Food Science and Engineering, South China University of TechnologySchool of Food Science and Engineering, South China University of TechnologyAbstract 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 Abstracthttps://doi.org/10.1186/s13068-022-02186-yRecombinant collagenasesSaccharomyces cerevisiaeHeterologous expressionCollagen degradationSynergistic effect |
| spellingShingle | Han Xiao Xiufang Liu Yunzi Feng Lin Zheng Mouming Zhao Mingtao Huang Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation Biotechnology for Biofuels and Bioproducts Recombinant collagenases Saccharomyces cerevisiae Heterologous expression Collagen degradation Synergistic effect |
| title | Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation |
| title_full | Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation |
| title_fullStr | Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation |
| title_full_unstemmed | Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation |
| title_short | Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation |
| title_sort | secretion of collagenases by saccharomyces cerevisiae for collagen degradation |
| topic | Recombinant collagenases Saccharomyces cerevisiae Heterologous expression Collagen degradation Synergistic effect |
| url | https://doi.org/10.1186/s13068-022-02186-y |
| work_keys_str_mv | AT hanxiao secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation AT xiufangliu secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation AT yunzifeng secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation AT linzheng secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation AT moumingzhao secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation AT mingtaohuang secretionofcollagenasesbysaccharomycescerevisiaeforcollagendegradation |