{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,5]],"date-time":"2026-04-05T22:24:12Z","timestamp":1775427852473,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,3,26]],"date-time":"2023-03-26T00:00:00Z","timestamp":1679788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2018YFA09013400"],"award-info":[{"award-number":["2018YFA09013400"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["21908239"],"award-info":[{"award-number":["21908239"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["TSBICIP-PTJS-001"],"award-info":[{"award-number":["TSBICIP-PTJS-001"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Natural Science Foundation of China","award":["2018YFA09013400"],"award-info":[{"award-number":["2018YFA09013400"]}]},{"name":"National Natural Science Foundation of China","award":["21908239"],"award-info":[{"award-number":["21908239"]}]},{"name":"National Natural Science Foundation of China","award":["TSBICIP-PTJS-001"],"award-info":[{"award-number":["TSBICIP-PTJS-001"]}]},{"name":"Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project","award":["2018YFA09013400"],"award-info":[{"award-number":["2018YFA09013400"]}]},{"name":"Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project","award":["21908239"],"award-info":[{"award-number":["21908239"]}]},{"name":"Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project","award":["TSBICIP-PTJS-001"],"award-info":[{"award-number":["TSBICIP-PTJS-001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>The naturally occurring one-carbon assimilation pathways for the production of acetyl-CoA and its derivatives often have low product yields because of carbon loss as CO2. We constructed a methanol assimilation pathway to produce poly-3-hydroxybutyrate (P3HB) using the MCC pathway, which included the ribulose monophosphate (RuMP) pathway for methanol assimilation and non-oxidative glycolysis (NOG) for acetyl-CoA (precursor for PHB synthesis) production. The theoretical product carbon yield of the new pathway is 100%, hence no carbon loss. We constructed this pathway in E. coli JM109 by introducing methanol dehydrogenase (Mdh), a fused Hps\u2013phi (hexulose-6-phosphate synthase and 3-phospho-6-hexuloisomerase), phosphoketolase, and the genes for PHB synthesis. We also knocked out the frmA gene (encoding formaldehyde dehydrogenase) to prevent the dehydrogenation of formaldehyde to formate. Mdh is the primary rate-limiting enzyme in methanol uptake; thus, we compared the activities of three Mdhs in vitro and in vivo and then selected the one from Bacillus methanolicus MGA3 for further study. Experimental results indicate that, in agreement with the computational analysis results, the introduction of the NOG pathway is essential for improving PHB production (65% increase in PHB concentration, up to 6.19% of dry cell weight). We demonstrated that PHB can be produced from methanol via metabolic engineering, which provides the foundation for the future large-scale use of one-carbon compounds for biopolymer production.<\/jats:p>","DOI":"10.3390\/bioengineering10040415","type":"journal-article","created":{"date-parts":[[2023,3,27]],"date-time":"2023-03-27T03:01:14Z","timestamp":1679886074000},"page":"415","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Engineering Escherichia coli for Poly-\u03b2-hydroxybutyrate Production from Methanol"],"prefix":"10.3390","volume":"10","author":[{"given":"Jiaying","family":"Wang","sequence":"first","affiliation":[{"name":"Tianjin University of Science and Technology, Tianjin 300457, China"},{"name":"Biodesign Center, Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"College of Food Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0027-2770","authenticated-orcid":false,"given":"Zhiqiang","family":"Chen","sequence":"additional","affiliation":[{"name":"Biodesign Center, Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaogui","family":"Deng","sequence":"additional","affiliation":[{"name":"Tianjin University of Science and Technology, Tianjin 300457, China"},{"name":"Biodesign Center, Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"School of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qianqian","family":"Yuan","sequence":"additional","affiliation":[{"name":"Biodesign Center, Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5325-2314","authenticated-orcid":false,"given":"Hongwu","family":"Ma","sequence":"additional","affiliation":[{"name":"Biodesign Center, Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"124550","DOI":"10.1016\/j.biortech.2020.124550","article-title":"Integrating anaerobic digestion of potato peels to methanol production by methanotrophs immobilized on banana leaves","volume":"323","author":"Patel","year":"2021","journal-title":"Bioresour. 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