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Engineering of two thiamine diphosphate-dependent enzymes for the regioselective condensation of C1-formaldehyde into C4-erythrulose.
Kim, Jun-Hong; Cheon, Huijin; Jo, Hye-Jin; Kim, Ji-Won; Kim, Ga Young; Seo, Hye-Rim; Seo, Pil-Won; Kim, Jeong-Sun; Park, Jin-Byung.
Afiliación
  • Kim JH; Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Cheon H; Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
  • Jo HJ; Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
  • Kim JW; Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Kim GY; Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
  • Seo HR; Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
  • Seo PW; Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Kim JS; Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: jsunkim@chonnam.ac.kr.
  • Park JB; Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea. Electronic address: jbpark06@ewha.ac.kr.
Int J Biol Macromol ; 253(Pt 8): 127674, 2023 Dec 31.
Article en En | MEDLINE | ID: mdl-37890751
A number of carboligases, which catalyze condensation of C1- and/or C2-aldehydes into multi-carbon products, have been reported. However, their catalytic activities and/or regioselectivities remained rather low. Thereby, this study has focused on engineering of C1 and C2 carboligases for the regioselective condensation of C1-formaldehyde into C4-erythrulose via C2-glycolaldehyde. The crystal structure of the glyoxylate carboligase from Escherichia coli (EcGCL) was elucidated in complex with glycolaldehyde. A structure-guided rationale generated several mutants, one of whose catalytic activity reached 15.6 M-1·s-1, almost 10 times greater than the wild-type enzyme. Another variant (i.e., EcGCL_R484M/N283Q/L478M/M488L/R284K) has shown significantly increased stability to the glycolaldehyde toxicity, enabling production of glycolaldehyde to 31 mM from 75 mM formaldehyde (conversion: 83 %). Besides, the E1 subunit of α-ketoglutarate dehydrogenase complex from Vibrio vulnificus (VvSucA) was engineered as a regiospecific C2 carboligase for condensation of glycolaldehyde into erythrulose. The combination of EcGCL_R484M/N283Q/L478M/M488L/R284K and VvSucA_K228L led to the cascade production of erythrulose to 8 mM from 90 mM formaldehyde via glycolaldehyde without byproduct formation. This study will contribute to valorization of C1 gases into industrially relevant multi-carbon products in an environment-friendly way.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Tiamina Pirofosfato / Escherichia coli Idioma: En Revista: Int J Biol Macromol Año: 2023 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
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Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Tiamina Pirofosfato / Escherichia coli Idioma: En Revista: Int J Biol Macromol Año: 2023 Tipo del documento: Article Pais de publicación: Países Bajos