{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T09:19:27Z","timestamp":1770974367848,"version":"3.50.1"},"reference-count":29,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T00:00:00Z","timestamp":1679443200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2023,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>R\u20101,3\u2010butanediol (R\u20101,3\u2010BDO) is an important chiral intermediate of penem and carbapenem synthesis. Among the different synthesis methods to obtain pure enantiomer R\u20101,3\u2010BDO, oxidation\u2013reduction cascades catalysed by enzymes are promising strategies for its production. Dehydrogenases have been used for the reduction step, but the enantio\u2010selectivity is not high enough for further organic synthesis efforts. Here, a short\u2010chain carbonyl reductase (LnRCR) was evaluated for the reduction step and developed via protein engineering. After docking result analysis with the substrate 4\u2010hydroxy\u20102\u2010butanone (4H2B), residues were selected for virtual mutagenesis, their substrate\u2010binding energies were compared, and four sites were selected for saturation mutagenesis. High\u2010throughput screening helped identify a Ser154Lys mutant which increased the catalytic efficiency by 115% compared to the parent enzyme. Computer\u2010aided simulations indicated that after single residue replacement, movements in two flexible areas (VTDPAF and SVGFANK) facilitated the volumetric compression of the 4H2B\u2010binding pocket. The number of hydrogen bonds between the stabilized 4H2B\u2010binding pocket of the mutant enzyme and substrate was higher (from four to six) than the wild\u2010type enzyme, while the substrate\u2010binding energy was decreased (from \u221217.0\u00a0kJ\/mol to \u221229.1\u00a0kJ\/mol). Consequently, the catalytic efficiency increased by approximately 115% and enantio\u2010selectivity increased from 95% to 99%. Our findings indicate that compact and stable substrate\u2010binding pockets are critical for enzyme catalysis. Lastly, the utilization of a microbe expressing the Ser154Lys mutant enzyme was proven to be a robust process to conduct the oxidation\u2013reduction cascade at larger scales.<\/jats:p>","DOI":"10.1111\/1751-7915.14249","type":"journal-article","created":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T06:21:21Z","timestamp":1679466081000},"page":"1333-1343","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A short\u2010chain carbonyl reductase mutant is an efficient catalyst in the production of (R)\u20101,3\u2010butanediol"],"prefix":"10.1111","volume":"16","author":[{"given":"Xiaoyan","family":"Guo","sequence":"first","affiliation":[{"name":"College of New Materials and Chemical Engineering Beijing Institute of Petrochemical Technology  Beijing China"},{"name":"Beijing Key Laboratory of Fuels Cleaning and Advanced Catalytic Emission Reduction Technology  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunfang","family":"Gao","sequence":"additional","affiliation":[{"name":"College of New Materials and Chemical Engineering Beijing Institute of Petrochemical Technology  Beijing China"},{"name":"Beijing Key Laboratory of Fuels Cleaning and Advanced Catalytic Emission Reduction Technology  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fangzheng","family":"Liu","sequence":"additional","affiliation":[{"name":"College of New Materials and Chemical Engineering Beijing Institute of Petrochemical Technology  Beijing China"},{"name":"Beijing Key Laboratory of Fuels Cleaning and Advanced Catalytic Emission Reduction Technology  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong","family":"Tao","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Microbial Physiological and Metabolic Engineering Institute of Microbiology, Chinese Academy of Sciences  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haibo","family":"Jin","sequence":"additional","affiliation":[{"name":"College of New Materials and Chemical Engineering Beijing Institute of Petrochemical Technology  Beijing China"},{"name":"Beijing Key Laboratory of Fuels Cleaning and Advanced Catalytic Emission Reduction Technology  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8205-7753","authenticated-orcid":false,"given":"Jianjun","family":"Wang","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Microbial Physiological and Metabolic Engineering Institute of Microbiology, Chinese Academy of Sciences  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheng","family":"Wu","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Microbial Physiological and Metabolic Engineering Institute of Microbiology, Chinese Academy of 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