PMID: 11917150Mar 28, 2002Paper

Engineering of multiple arginines into the Ser/Thr surface of Trichoderma reesei endo-1,4-beta-xylanase II increases the thermotolerance and shifts the pH optimum towards alkaline pH

Protein Engineering
Ossi TurunenM Leisola

Abstract

We studied the effects of increase in the number of surface arginines on the enzyme activity and stability of Trichoderma reesei endo-1,4-beta-xylanase II. The number of arginines was increased in two mutant series. The first set contained six arginines on different sides of the protein surface. These arginines had no significant effect on the thermostability. However, the optimal pH region became narrower. Another series of five arginines was engineered into the 'Ser/Thr surface', formed of part of the double-layered beta-sheet located on one side of the 'right-hand-like' xylanase. These mutations shifted the activity profile to the alkaline region by approximately 0.5-1.0 pH units. In addition, the arginines on the Ser/Thr surface increased the enzyme activity at high temperature, although the enzyme stability in the absence of substrate decreased significantly at 50-55 degrees C. In the presence of the substrate, the thermostability increased 4-5-fold at 60-65 degrees C. Thus, the substrate neutralized the destabilizing effect of Ser/Thr surface arginines and revealed a stabilizing effect of the same mutations. The stabilizing effect of arginines at high temperatures was seen clearly only when five arginines were introduced ...Continue Reading

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Citations

Aug 31, 2012·Bioprocess and Biosystems Engineering·Vikash KumarT Satyanarayana
Jul 28, 2004·Extremophiles : Life Under Extreme Conditions·Hairong XiongOssi Turunen
Aug 21, 2013·Applied Biochemistry and Biotechnology·Shunchen Qiu, Luhua Lai
Jun 29, 2011·Protein Engineering, Design & Selection : PEDS·Craig A HokansonRoberto Crea
Aug 24, 2005·Bioscience, Biotechnology, and Biochemistry·Hajime ShibuyaKiyoshi Hayashi
Aug 7, 2014·Protein Engineering, Design & Selection : PEDS·Roberto RullerRichard John Ward
Jan 13, 2016·World Journal of Microbiology & Biotechnology·Vishal KumarPratyoosh Shukla
Sep 11, 2004·Biochemical and Biophysical Research Communications·Liangwei LiuWeilan Shao
Jan 18, 2005·FEMS Microbiology Reviews·Tony CollinsGeorges Feller
Feb 18, 2009·FEMS Microbiology Letters·Dawn Elizabeth StephensKugen Permaul
May 4, 2016·Applied Microbiology and Biotechnology·Tomohiko MatsuzawaKatsuro Yaoi
Aug 9, 2006·Journal of Biotechnology·Dawn Elizabeth StephensSuren Singh
Dec 14, 2007·Journal of Biotechnology·Ji Hye YangYoung Je Yoo
Oct 25, 2017·Applied Microbiology and Biotechnology·Tomohiko MatsuzawaKatsuro Yaoi
Apr 4, 2008·Proteins·Gwénaëlle André-LerouxNathalie Juge

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