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Enzymatic properties of alcohol dehydrogenase PedE_M.s. derived from Methylopila sp. M107 and its broad metal selectivity.


ABSTRACT: As an important metabolic enzyme in methylotrophs, pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenases play significant roles in the global carbon and nitrogen cycles. In this article, a calcium (Ca2+)-dependent alcohol dehydrogenase PedE_M.s., derived from the methylotroph Methylopila sp. M107 was inserted into the modified vector pCM80 and heterologously expressed in the host Methylorubrum extorquens AM1. Based on sequence analysis, PedE_M.s., a PQQ-dependent dehydrogenase belonging to a methanol/ethanol family, was successfully extracted and purified. Showing by biochemical results, its enzymatic activity was detected as 0.72 U/mg while the Km value was 0.028 mM while employing ethanol as optimal substrate. The activity of PedE_M.s. could be enhanced by the presence of potassium (K+) and calcium (Ca2+), while acetonitrile and certain common detergents have been found to decrease the activity of PedE_M.s.. In addition, its optimum temperature and pH were 30°C and pH 9.0, respectively. Chiefly, as a type of Ca2+-dependent alcohol dehydrogenase, PedE_M.s. maintained 60-80% activity in the presence of 10 mM lanthanides and displayed high affinity for ethanol compared to other PedE-type enzymes. The 3D structure of PedE_M.s. was predicted by AlphaFold, and it had an 8-bladed propeller-like super-barrel. Meanwhile, we could speculate that PedE_M.s. contained the conserved residues Glu213, Asn300, and Asp350 through multiple sequence alignment by Clustal and ESpript. The analysis of enzymatic properties of PedE_M.s. enriches our knowledge of the methanol/ethanol family PQQ-dependent dehydrogenase. This study provides new ideas to broaden the application of alcohol dehydrogenase in alcohol concentration calculation, biosensor preparation, and other industries.

SUBMITTER: Xiao Y 

PROVIDER: S-EPMC10409515 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature

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Enzymatic properties of alcohol dehydrogenase PedE_M.s. derived from <i>Methylopila</i> sp. M107 and its broad metal selectivity.

Xiao Ying Y   Wu Kaijuan K   Batool Syeda Sundas SS   Wang Qingqun Q   Chen Hao H   Zhai Xingyu X   Yu Zheng Z   Huang Jing J  

Frontiers in microbiology 20230725


As an important metabolic enzyme in methylotrophs, pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenases play significant roles in the global carbon and nitrogen cycles. In this article, a calcium (Ca<sup>2+</sup>)-dependent alcohol dehydrogenase PedE_M.s., derived from the methylotroph <i>Methylopila</i> sp. M107 was inserted into the modified vector pCM80 and heterologously expressed in the host <i>Methylorubrum extorquens</i> AM1. Based on sequence analysis, PedE_M.s., a PQQ-depende  ...[more]

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