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Molecular cloning of rhodanese gene from soil metagenome of cold desert of North-West Himalayas: sequence and structural features of the rhodanese enzyme.

ABSTRACT: Rhodanese is a multifunctional, sulfur transferase that catalyzes the detoxification of cyanide by sulphuration in a double displacement (ping pong) mechanistic reaction. In the present study, small-insert metagenomic library from soil sample collected from Ladakh (3,000-3,600 m.a.s.l) in northwestern Himalayas, India was constructed. Function-driven screening of ~8,500 colonies led to the isolation of one esterase-positive clone (clone-est) harboring 2.43 kb insert. Sequence analysis of the insert identified two ORF's, phosM encoding phosphoesterase and rodM encoding rhodanese. The 800 bp rodM gene encoded a polypeptide of 227 amino acids (RodM). The RodM showed maximum homology with the rhodanese-like protein from Cyanobacterium synechococcus species with a score identity of only 51 %. Putative 3D structure of RodM developed by homology modeling resembles to homodimeric protein of SUD sulfur transferase of Wolinella succinogenes with properly structured active-site cysteine (Cys) residue. Rhodanese has been reported from few culturable microorganisms.

SUBMITTER: Bhat A

PROVIDER: S-EPMC4522728 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Chickpea is one of the most important grain legume crops in the world. India is the largest producer, consumer as well as importer of chickpea. Cold stress (temperature < 15 °C) is one of the important abiotic stresses limiting chickpea production by hampering its growth and vigor at all phenological stages. This study was aimed to characterize a diverse set of 366 chickpea genotypes for cold tolerance and identify most promising cold tolerant chickpea genotypes in the Western-Himalayas of Jammu and Kashmir, India. The 366 genotypes used during the present study including genotypes belonging to cultivated, primary and secondary gene pools of chickpea. Two important approaches were used including visual screening under field conditions and screening under controlled conditions by measuring cell membrane stability through electrolyte leakage tests. The analysis of trait data collected through both the approaches led to the identification of five most promising/candidate cold tolerant chickpea genotypes including one wild genotype "Ortan-066" from secondary gene pool species (C. echinospermum), one wild genotype "Cudi 1-022" from primary gene pool species (C. reticulatum) and three genotypes (IC 116783, ICC 15200 and AGBLG 170004) from the cultivated species (Cicer arietinum). Wild genotype "Ortan-066" was found best cold tolerance source with the mean Cold Tolerance Rating (CTR) of 2 and Electrolyte Leakage Index (ELI) of 10.82%, followed by wild genotype "Cudi 1-022" (CTR = 3, ELI = 18.89%), and three cultivated genotypes viz., IC 116783, ICC 15200 and AGBL-G-170004, with the mean CTR of 3 and an estimated mean ELI of 21.26%, 21.58% and 21.94%, respectively. The promising, candidate cold tolerant genotypes identified during the present study could be used in chickpea breeding programs aimed at improving cold tolerance of cultivated chickpea worldwide. The candidate lines can be also used for developing bi-parental mapping populations, wild × cultivated introgression lines, transcriptomics and for differential expression analysis of cold tolerant genes in chickpea.Supplementary informationThe online version contains supplementary material available at 10.1007/s12298-021-00997-1.

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Molecular cloning of rhodanese gene from soil metagenome of cold desert of North-West Himalayas: sequence and structural features of the rhodanese enzyme.

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