Analysis of the raw starch-binding domain by mutation of a glucoamylase from Aspergillus awamori var. kawachi expressed in Saccharomyces cerevisiae.

Applied and Environmental Microbiology
Masatoshi GotoShinsaku Hayashida

Abstract

Carboxy-terminal deletions were introduced into the raw starch-binding domain (A-515 to R-615) encoded by the gene for glucoamylase I (GAI) from Aspergillus awamori var. kawachi. Genes coding for proteins designated GA596 (A-1 to E-596), GA570 (A-1 to A-570), and GA559 (A-1 to N-559) were constructed and resulted in truncated proteins. All of the mutant genes were expressed heterologously in Saccharomyces cerevisiae. GA596 adsorbed to raw starch and digested it. GA570 and GA559 did not adsorb to raw starch or to an alpha-cyclodextrin-Sepharose CL-4B gel under our experimental conditions. However, GA570 was able to digest raw starch, and the digestion of raw starch by GA570 was inhibited by beta-cyclodextrin. Residue Trp-562 of GAI, which was suggested previously to contribute to formation of an inclusion complex with raw starch, was replaced by Leu (GAW562L), Phe (GAW562F), and Gly (GAW562G). GAW562L and GAW562F adsorbed to raw starch and an alpha-cyclodextrin gel, but GAW562G did not. Although GAW562L digested raw starch to the same extent as wild-type GAI (designated GAY), GAW562F and GAW562G exhibited less ability to digest raw starch. On the basis of our results, it appears that the sequence around Trp-562, PL(W-562)YVTVTLP...Continue Reading

References

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Citations

Mar 12, 2009·Applied Biochemistry and Biotechnology·Haiyan SunMing Peng
Mar 22, 2012·Applied Biochemistry and Biotechnology·Hui ZhaoTatsuzo Oka
Oct 31, 2002·Applied and Environmental Microbiology·Keisuke EkinoKensuke Furukawa
May 23, 2002·FEMS Microbiology Letters·Fabrice SabathéPhilippe Soucaille
Jan 1, 1996·Annual Review of Microbiology·R A Warren
Nov 23, 2006·The Biochemical Journal·Yu-Nan LiuPing-Chiang Lyu
Feb 19, 2008·FEMS Microbiology Letters·Alexsandro S GaldinoFernando A G Torres
Jun 26, 2007·Bioscience, Biotechnology, and Biochemistry·Masatoshi Goto
Sep 8, 2010·Applied Biochemistry and Biotechnology·Ming-Chung ChengPao-Chuan Hsieh
Aug 18, 2009·The FEBS Journal·Camilla ChristiansenBirte Svensson
Dec 17, 2014·Applied and Environmental Microbiology·Taiki FutagamiMasatoshi Goto
Nov 13, 2009·Applied Microbiology and Biotechnology·Daniel GuillénRomina Rodríguez-Sanoja
May 27, 2017·Proteins·Silvia ArmentaRomina Rodríguez-Sanoja
Apr 13, 2018·Applied Biochemistry and Biotechnology·Tahmina HossainAbdullah-Al- Mahin
Aug 29, 2019·Proceedings of the National Academy of Sciences of the United States of America·Alan K ItakuraMartin C Jonikas
May 4, 2000·Enzyme and Microbial Technology·L M HamiltonW M Fogarty
Jun 9, 2005·Current Opinion in Microbiology·Romina Rodríguez-SanojaSergio Sánchez

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