Partial Gene Cloning and Enzyme Structure Modeling of Exolevanase Fragment from Bacillus subtilis

2nd International Conference on Mathematics, Science, Education and Technology

数学;自然科学;教育

Azhar, M.^1 ; Natalia, D.^2 ; Syukur, S.^3 ; Andriani, N.^1 ; Jamsari, J.^4

Biochemistry Laboratory, Faculty of Mathematics and Natural Sciences, Universitas Negeri Padang, Jl. Prof. Hamka, Air Tawar, Padang

25131, Indonesia^1

Biochemistry Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha10, Bandung

40132, Indonesia^2

Biochemistry Laboratory, Faculty of Mathematics and Natural Sciences, Universitas Andalas, Padang

25163, Indonesia^3

Biotechnology Laboratory, Faculty of Agriculture, Universitas Andalas, Padang

25163, Indonesia^4

关键词: Bacillus Subtilis;    Enzyme structures;    Inulin hydrolysis;    Polymerase chain reaction methods;    Potential sources;    Protein fragments;    Substrate binding;    Thermotolerant;   

Others  :  https://iopscience.iop.org/article/10.1088/1757-899X/335/1/012026/pdf
DOI  :  10.1088/1757-899X/335/1/012026

来源: IOP

【 摘 要 】

Inulin hydrolysis thermophilic and thermotolerant bacteria are potential sources of inulin hydrolysis enzymes. Partial gene that encodes inulin hydrolysis enzymes had been isolated from Bacillus subtilis using polymerase chain reaction (PCR) method with the DPE.slFandDPE.eR degenerative primers. The partial gene was cloned into pGEM-T Easy vector with E. coli as host cells and analyzed using BLASTx, CrustalW2, and Phyre2 programs. Size of thepartial gene had been found539 bp that encoded 179aminoacid residues of protein fragment. The sequences of protein fragment was more similar to exolevanase than exoinulinase. The protein fragment had conserved motif FSGS, and specific hits GH32 β-fructosidase. It had three residues of active site and five residues of substrate binding. The active site on the protein fragment were D (1-WLNDP-5), D (125-FRDPK-129) and E (177-WEC-179). Substrate binding on the protein fragment were ND (1-WLNDP-5), Q (18-FYQY-21), FS (60-FSGS-63) RD (125-FRDPK-129) and E (177-WEC-179).