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Isolation of a Bacillus cereus strain HBL‐AI and its application for production of T rans ‐4‐hydroxy‐ l ‐proline
Author(s) -
Wang X.M.,
Han M.N.,
Jiang J.P.,
Fu S.Q.,
Zhang F.H.,
Du J.,
Zhang H.L.,
Li W.
Publication year - 2021
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.13388
Subject(s) - bacillus cereus , proline , escherichia coli , biochemistry , bioconversion , enzyme , biology , bacteria , strain (injury) , metabolic engineering , chemistry , gene , amino acid , fermentation , genetics , anatomy
A new trans ‐4‐hydroxy‐ l ‐proline ( trans ‐Hyp) producing Bacillus cereus HBL‐AI, was isolated from the air, which was screened just using l ‐proline as carbon and energy sources. This strain exhibited 73·4% bioconversion rate from initial l ‐proline (3 g l −1 ) to trans ‐Hyp. By sequencing the genome of this bacterium, 6244 coding sequences were obtained. Genome annotation analysis and functional expression were used to identify the proline‐4‐hydroxylase (BP4H) in HBL‐AI. This enzyme belonged to a family of 2‐oxoglutarate‐related dioxygenases, which required 2‐oxoglutarate and O 2 as co‐substrates for the reaction. Homologous modelling indicated that the enzyme had two monomers and contained conserved motifs, which included a distorted ‘jelly roll’ β strand core and the residues (HXDXnH and RXS). The engineering Escherichia coli 3 Δ W3110/ pTrc99a‐proba‐bp4h was constructed using BP4H, which transformed glucose to trans ‐Hyp in one step with high concentration of 46·2 g l −1 . This strategy provides a green and efficient method for synthesis of trans ‐Hyp and thus has a great potential in industrial application.