Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase
Fatty acid ethyl esters are important flavor chemicals in strong-flavor Baijiu. Monascus purpureus YJX-8 is recognized as an important microorganism for ester synthesis in the fermentation process. Enzyme LIP05 from YJX-8 can efficiently catalyze the synthesis of fatty acid ethyl esters under aqueou...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-01-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1107104/full |
| _version_ | 1828066520943558656 |
|---|---|
| author | Jingrong Zhao Jingrong Zhao Youqiang Xu Youqiang Xu Hongyun Lu Hongyun Lu Dong Zhao Jia Zheng Mengwei Lin Mengwei Lin Xin Liang Xin Liang Ze Ding Ze Ding Wenqi Dong Wenqi Dong Maochen Yang Weiwei Li Weiwei Li Chengnan Zhang Chengnan Zhang Baoguo Sun Baoguo Sun Xiuting Li Xiuting Li Xiuting Li |
| author_facet | Jingrong Zhao Jingrong Zhao Youqiang Xu Youqiang Xu Hongyun Lu Hongyun Lu Dong Zhao Jia Zheng Mengwei Lin Mengwei Lin Xin Liang Xin Liang Ze Ding Ze Ding Wenqi Dong Wenqi Dong Maochen Yang Weiwei Li Weiwei Li Chengnan Zhang Chengnan Zhang Baoguo Sun Baoguo Sun Xiuting Li Xiuting Li Xiuting Li |
| author_sort | Jingrong Zhao |
| collection | DOAJ |
| description | Fatty acid ethyl esters are important flavor chemicals in strong-flavor Baijiu. Monascus purpureus YJX-8 is recognized as an important microorganism for ester synthesis in the fermentation process. Enzyme LIP05 from YJX-8 can efficiently catalyze the synthesis of fatty acid ethyl esters under aqueous phase, but the key catalytic sites affecting esterification were unclear. The present work combined homology modeling, molecular dynamics simulation, molecular docking and site-directed mutation to analyze the catalytic mechanism of LIP05. Protein structure modeling indicated LIP05 belonged to α/β fold hydrolase, contained a lid domain and a core catalytic pocket with conserved catalytic triad Ser150-His215-Asp202, and the oxyanion hole composed of Gly73 and Thr74. Ile30 and Leu37 of the lid domain were found to affect substrate specificity. The π-bond stacking between Tyr116 and Tyr149 played an important role in stabilizing the catalytic active center of LIP05. Tyr116 and Ile204 determined the substrate spectrum by composing the substrate-entrance channel. Residues Leu83, Ile204, Ile211 and Leu216 were involved in forming the hydrophobic substrate-binding pocket through steric hindrance and hydrophobic interaction. The catalytic mechanism for esterification in aqueous phase of LIP05 was proposed and provided a reference for clarifying the synthesis of fatty acid ethyl esters during the fermentation process of strong-flavor Baijiu. |
| first_indexed | 2024-04-10T23:31:55Z |
| format | Article |
| id | doaj.art-8f56c886e1d8449389a4665ac8be0abb |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-10T23:31:55Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-8f56c886e1d8449389a4665ac8be0abb2023-01-12T05:10:41ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-01-011310.3389/fmicb.2022.11071041107104Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phaseJingrong Zhao0Jingrong Zhao1Youqiang Xu2Youqiang Xu3Hongyun Lu4Hongyun Lu5Dong Zhao6Jia Zheng7Mengwei Lin8Mengwei Lin9Xin Liang10Xin Liang11Ze Ding12Ze Ding13Wenqi Dong14Wenqi Dong15Maochen Yang16Weiwei Li17Weiwei Li18Chengnan Zhang19Chengnan Zhang20Baoguo Sun21Baoguo Sun22Xiuting Li23Xiuting Li24Xiuting Li25Key Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaWuliangye Yibin Co., Ltd., Yibin, Sichuan, ChinaWuliangye Yibin Co., Ltd., Yibin, Sichuan, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaSchool of Food and Health, Beijing Technology and Business University (BTBU), Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaSchool of Food and Health, Beijing Technology and Business University (BTBU), Beijing, ChinaSchool of Food and Health, Beijing Technology and Business University (BTBU), Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaKey Laboratory of Brewing Microbiome and Enzymatic Molecular Engineering, China General Chamber of Commerce, Beijing Technology and Business University, Beijing, ChinaBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, ChinaSchool of Food and Health, Beijing Technology and Business University (BTBU), Beijing, ChinaFatty acid ethyl esters are important flavor chemicals in strong-flavor Baijiu. Monascus purpureus YJX-8 is recognized as an important microorganism for ester synthesis in the fermentation process. Enzyme LIP05 from YJX-8 can efficiently catalyze the synthesis of fatty acid ethyl esters under aqueous phase, but the key catalytic sites affecting esterification were unclear. The present work combined homology modeling, molecular dynamics simulation, molecular docking and site-directed mutation to analyze the catalytic mechanism of LIP05. Protein structure modeling indicated LIP05 belonged to α/β fold hydrolase, contained a lid domain and a core catalytic pocket with conserved catalytic triad Ser150-His215-Asp202, and the oxyanion hole composed of Gly73 and Thr74. Ile30 and Leu37 of the lid domain were found to affect substrate specificity. The π-bond stacking between Tyr116 and Tyr149 played an important role in stabilizing the catalytic active center of LIP05. Tyr116 and Ile204 determined the substrate spectrum by composing the substrate-entrance channel. Residues Leu83, Ile204, Ile211 and Leu216 were involved in forming the hydrophobic substrate-binding pocket through steric hindrance and hydrophobic interaction. The catalytic mechanism for esterification in aqueous phase of LIP05 was proposed and provided a reference for clarifying the synthesis of fatty acid ethyl esters during the fermentation process of strong-flavor Baijiu.https://www.frontiersin.org/articles/10.3389/fmicb.2022.1107104/fullstrong-flavor Baijiufatty acid ethyl esterenzymatic esterificationaqueous phasemolecular mechanism |
| spellingShingle | Jingrong Zhao Jingrong Zhao Youqiang Xu Youqiang Xu Hongyun Lu Hongyun Lu Dong Zhao Jia Zheng Mengwei Lin Mengwei Lin Xin Liang Xin Liang Ze Ding Ze Ding Wenqi Dong Wenqi Dong Maochen Yang Weiwei Li Weiwei Li Chengnan Zhang Chengnan Zhang Baoguo Sun Baoguo Sun Xiuting Li Xiuting Li Xiuting Li Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase Frontiers in Microbiology strong-flavor Baijiu fatty acid ethyl ester enzymatic esterification aqueous phase molecular mechanism |
| title | Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase |
| title_full | Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase |
| title_fullStr | Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase |
| title_full_unstemmed | Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase |
| title_short | Molecular mechanism of LIP05 derived from Monascus purpureus YJX-8 for synthesizing fatty acid ethyl esters under aqueous phase |
| title_sort | molecular mechanism of lip05 derived from monascus purpureus yjx 8 for synthesizing fatty acid ethyl esters under aqueous phase |
| topic | strong-flavor Baijiu fatty acid ethyl ester enzymatic esterification aqueous phase molecular mechanism |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1107104/full |
| work_keys_str_mv | AT jingrongzhao molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT jingrongzhao molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT youqiangxu molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT youqiangxu molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT hongyunlu molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT hongyunlu molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT dongzhao molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT jiazheng molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT mengweilin molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT mengweilin molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT xinliang molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT xinliang molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT zeding molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT zeding molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT wenqidong molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT wenqidong molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT maochenyang molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT weiweili molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT weiweili molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT chengnanzhang molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT chengnanzhang molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT baoguosun molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT baoguosun molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT xiutingli molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT xiutingli molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase AT xiutingli molecularmechanismoflip05derivedfrommonascuspurpureusyjx8forsynthesizingfattyacidethylestersunderaqueousphase |