Cloning, Expression and Enzymatic Properties Analysis of Dihydrofolate Reductase Gene from the Silkworm, Bombyx Mori

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2012 Oct 16

PMID 23065260

Citations 2

Authors

Wenjing Wang

Junshan Gao

Jing Wang

Chaoliang Liu

Yan Meng

Affiliations

Soon will be listed here.

Abstract

Tetrahydrobiopterin (BH4) is an essential cofactor for aromatic acid hydroxylases, which control the levels of monoamine neurotransmitters. BH4 deficiency has been associated with many neuropsychological disorders. Dihydrofolate reductase (DHFR) can catalyze 7,8-dihydrobiopterin to 5,6,7,8-tetrahydrobiopterin (BH4) in the salvage pathway of BH4 synthesis from sepiapterin (SP), a major pigment component contained in the integument of silkworm Bombyx mori mutant lemon (lem) in high concentration. In this study, we report the cloning of DHFR gene from the silkworm B. mori (BmDhfr) and identification of enzymatic properties of BmDHFR. BmDhfr is located on scaffold Bm_199 with a predicted gene model BGIBMGA013340, which encodes a 185-aa polypeptide with a predicted molecular mass of about 21 kDa. Biochemical analyses showed that the recombinant BmDHFR protein exhibited high enzymatic activity and suitable parameters to substrate. Together with our previous studies on SP reductase of B. mori (BmSPR) and the lem mutant, it may be an effective way to industrially extract SP from the lem silkworms in large scale to produce BH4 in vitro by co-expressing BmSPR and BmDHFR and using the extracted SP as a substrate in the future.

Citing Articles

Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus.

Song X, Hu D, Yan M, Wang Y, Wang N, Gu X Sci Rep. 2017; 7(1):514.

PMID: 28364125 PMC: 5428777. DOI: 10.1038/s41598-017-00643-5.

Molecular Characterization of QDPR Gene in Iranian Families with BH4 Deficiency: Reporting Novel and Recurrent Mutations.

Foroozani H, Abiri M, Salehpour S, Bagherian H, Sharifi Z, Alaei M JIMD Rep. 2015; 21:123-8.

PMID: 26006720 PMC: 4470949. DOI: 10.1007/8904_2015_441.

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