High-level expression of Cephalosporin C deacetylase from Bacillus subtilis SIL3 in Escherichia coli by a multilevel collaborative strategy

Ma, Xiaoqiang; Zhu, Ying; Sun, Ming; Yang, Shengli; Su, Erzheng; Wei, Dongzhi

doi:10.1016/j.bej.2016.07.009

Cited by 3 publications

(2 citation statements)

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“…Two major methods, namely, chemical and enzymatic deacetylation of 7-ACA, could be used to generate D-7-ACA. The enzymatic processes using deacetylases or esterases have attracted more attention and have gradually become the prevailing trend for the production of D-7-ACA, compared with the chemical approaches that require energy-intensive conditions and generate toxic waste ( Ma et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, exploring and developing new types of effective antibacterial antibiotics are urgent (Theuretzbacher et al, 2020). Deacetyl-7-aminocephalosporanic acid (D-7-ACA) is 10.3389/fmicb.2023.1164815 an important intermediate for synthesizing new kinds of semisynthetic cephalosporins due to the modification of the 3 hydroxyl group of D-7-ACA which is easy to be achieved (Tan et al, 2018), such as the generation of clinically used cefoxitin and cefuroxime (Ma et al, 2015(Ma et al, , 2016. Two major methods, namely, chemical and enzymatic deacetylation of 7-ACA, could be used to generate D-7-ACA.…”

Section: Introductionmentioning

confidence: 99%

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Biochemical characterization of an acetylesterase from Bacillus subtilis and its application for 7-aminocephalosporanic acid deacetylation

Wang,

Nong,

et al. 2023

Front. Microbiol.

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Deacetyl-7-aminocephalosporanic acid (D-7-ACA), which could be converted from 7-aminocephalosporanic acid (7-ACA), is a crucial starting material that is used for synthesizing industrial semisynthetic β-lactam antibiotics. Enzymes involved in the conversion from 7-ACA to D-7-ACA present critical resources in the pharmaceutical industry. In the present study, a putative acetylesterase, EstSJ, identified from Bacillus subtilis KATMIRA1933, was first heterologously expressed in Escherichia coli BL21(DE3) cells and biochemically characterized. EstSJ belongs to carbohydrate esterase family 12 and is active on short-chain acyl esters from p-NPC2 to p-NPC6. Multiple sequence alignments showed that EstSJ was also an SGNH family esterase with a typical GDS(X) motif at its N-terminal end and a catalytic triad composed of Ser186-Asp354-His357. The purified EstSJ displayed the highest specific activity of 1,783.52 U mg–1 at 30°C and pH 8.0, and was stable within the pH range of 5.0–11.0. EstSJ can deacetylate the C3′ acetyl group of 7-ACA to generate D-7-ACA, and the deacetylation activity was 4.50 U mg–1. Based on the structural and molecular docking with 7-ACA, the catalytic active sites (Ser186-Asp354-His357) together with four substrate-binding residues (Asn259, Arg295, Thr355, and Leu356) of EstSJ are revealed. This study provided a promising 7-ACA deacetylase candidate that could be applied to produce D-7-ACA from 7-ACA in the pharmaceutical industry.

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Section: Introductionmentioning

confidence: 99%