Impact of Leucine 278 Residue on Fatty Acid Length Specificity of Candida antarctica Lipase B - Advances in Microbiology

AiM > Vol.5 No.7, July 2015

Advances in Microbiology

Volume 5, Issue 7 (July 2015)

ISSN Print: 2165-3402 ISSN Online: 2165-3410

Google-based Impact Factor: 1.35 Citations

Impact of Leucine 278 Residue on Fatty Acid Length Specificity of Candida antarctica Lipase B ()

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DOI: 10.4236/aim.2015.57050 3,626 Downloads 5,068 Views Citations

Author(s)

Fanghua Wang¹, Shulin Hou², Qian Wang³, Pu Wang³, Jinsong Liu², Bo Yang³, Yonghua Wang^1*

Affiliation(s)

¹College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
²State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
³School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China.

ABSTRACT

Structural analysis of Candida antarctica lipase B (CALB) indicates that side chain of leucine at 278 site lies above the entrance of the catalytic pocket, which prognosticates its potential role on substrate specificity of the enzyme. To verify this presumption, shortened side chain of glycine or proline was rational designed and mutants were constructed by site-directed mutagenesis method. The colorimetric assay using p-nitrophenyl esters of fatty acids with various chain-lengths was used to study the substrate preference of lipases. Results indicated that L278G or L278P mutations both induced the drift of substrate specificity of CALB from p-nitrophenyl caprylate (pNP-C8) to longer carbon chain length of p-nitrophenyl caprate (pNP-C10). Meanwhile, V_max value of two mutants to pNP-C10 was both higher than that of wild-type. Docking results also indicated that shortened side chain of glycine or proline residues substitution at this site could get rid of the space block present above the catalytic pocket, and made longer chain substrate (pNP-C10) enter into the catalytic pocket easier. The modulation of specificity observed allowed for building substrate binding model and opened new possibilities for designing ligand specific lipases.

KEYWORDS

Candida antarctica Lipase B (CALB), Site-Directed Mutagenesis, Substrate Specificity, Stereospecific Blockade

Share and Cite:

Wang, F. , Hou, S. , Wang, Q. , Wang, P. , Liu, J. , Yang, B. and Wang, Y. (2015) Impact of Leucine 278 Residue on Fatty Acid Length Specificity of Candida antarctica Lipase B. Advances in Microbiology, 5, 493-499. doi: 10.4236/aim.2015.57050.

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