Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase

Tauna R. Rignall; John O. Baker; Suzanne L. McCarter; William S. Adney; Todd B. Vinzant; Stephen R. Decker; Michael E. Himmel

doi:10.1385/ABAB:98-100:1-9:383

Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase

Tauna R. Rignall, John O. Baker, Suzanne L. McCarter, William S. Adney, Todd B. Vinzant, Stephen R. Decker, Michael E. Himmel

National Renewable Energy Laboratory

Research output: Contribution to journal › Article › peer-review

27 Scopus Citations

Abstract

Mutation of a single active-site cleft tyrosyl residue to a glycyl residue significantly changes the mixture of products released from phosphoric acid-swollen cellulose (PSC) by EIcd, the catalytic domain of the endoglucanase-I from Acidothermus cellulolyticus. The percentage of glucose in the product stream is almost 40% greater for the Y245G mutant (and for an additional double mutant, Y245G/Q204A) than for the wild type enzyme. Comparisons of results for digestion PSC and of pretreated yellow poplar suggest that the observed shifts in product specificity are connected to the hydrolysis of a more easily digestible fraction of both substrates. A model is presented that relates the changes in product specificity to a mutation-driven shift in indexing of the polymeric substrate along the extended binding-site cleft.

Original language	American English
Pages (from-to)	383-394
Number of pages	12
Journal	Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
Volume	98-100
DOIs	https://doi.org/10.1385/ABAB:98-100:1-9:383 https://doi.org/10.1385/ABAB:98-100:1-9:383
State	Published - 2002

Bibliographical note

Posted with permission.

NREL Publication Number

NREL/JA-510-30264

Keywords

Acidothermus cellulolyticus
Cellulose
Endoglucanase
Product specificity
Saccharification

Access to Document

https://www.nrel.gov/docs/gen/fy02/30264.pdf

Cite this

Rignall, T. R., Baker, J. O., McCarter, S. L., Adney, W. S., Vinzant, T. B., Decker, S. R., & Himmel, M. E. (2002). Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase. Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, 98-100, 383-394. https://doi.org/10.1385/ABAB:98-100:1-9:383, https://doi.org/10.1385/ABAB:98-100:1-9:383

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title = "Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase",

abstract = "Mutation of a single active-site cleft tyrosyl residue to a glycyl residue significantly changes the mixture of products released from phosphoric acid-swollen cellulose (PSC) by EIcd, the catalytic domain of the endoglucanase-I from Acidothermus cellulolyticus. The percentage of glucose in the product stream is almost 40% greater for the Y245G mutant (and for an additional double mutant, Y245G/Q204A) than for the wild type enzyme. Comparisons of results for digestion PSC and of pretreated yellow poplar suggest that the observed shifts in product specificity are connected to the hydrolysis of a more easily digestible fraction of both substrates. A model is presented that relates the changes in product specificity to a mutation-driven shift in indexing of the polymeric substrate along the extended binding-site cleft.",

keywords = "Acidothermus cellulolyticus, Cellulose, Endoglucanase, Product specificity, Saccharification",

author = "Rignall, {Tauna R.} and Baker, {John O.} and McCarter, {Suzanne L.} and Adney, {William S.} and Vinzant, {Todd B.} and Decker, {Stephen R.} and Himmel, {Michael E.}",

note = "Posted with permission.",

year = "2002",

doi = "10.1385/ABAB:98-100:1-9:383",

language = "American English",

volume = "98-100",

pages = "383--394",

journal = "Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology",

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Rignall, TR, Baker, JO, McCarter, SL, Adney, WS, Vinzant, TB, Decker, SR & Himmel, ME 2002, 'Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase', Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, vol. 98-100, pp. 383-394. https://doi.org/10.1385/ABAB:98-100:1-9:383, https://doi.org/10.1385/ABAB:98-100:1-9:383

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T1 - Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase

AU - Rignall, Tauna R.

AU - Baker, John O.

AU - McCarter, Suzanne L.

AU - Adney, William S.

AU - Vinzant, Todd B.

AU - Decker, Stephen R.

AU - Himmel, Michael E.

N1 - Posted with permission.

PY - 2002

Y1 - 2002

N2 - Mutation of a single active-site cleft tyrosyl residue to a glycyl residue significantly changes the mixture of products released from phosphoric acid-swollen cellulose (PSC) by EIcd, the catalytic domain of the endoglucanase-I from Acidothermus cellulolyticus. The percentage of glucose in the product stream is almost 40% greater for the Y245G mutant (and for an additional double mutant, Y245G/Q204A) than for the wild type enzyme. Comparisons of results for digestion PSC and of pretreated yellow poplar suggest that the observed shifts in product specificity are connected to the hydrolysis of a more easily digestible fraction of both substrates. A model is presented that relates the changes in product specificity to a mutation-driven shift in indexing of the polymeric substrate along the extended binding-site cleft.

AB - Mutation of a single active-site cleft tyrosyl residue to a glycyl residue significantly changes the mixture of products released from phosphoric acid-swollen cellulose (PSC) by EIcd, the catalytic domain of the endoglucanase-I from Acidothermus cellulolyticus. The percentage of glucose in the product stream is almost 40% greater for the Y245G mutant (and for an additional double mutant, Y245G/Q204A) than for the wild type enzyme. Comparisons of results for digestion PSC and of pretreated yellow poplar suggest that the observed shifts in product specificity are connected to the hydrolysis of a more easily digestible fraction of both substrates. A model is presented that relates the changes in product specificity to a mutation-driven shift in indexing of the polymeric substrate along the extended binding-site cleft.

KW - Acidothermus cellulolyticus

KW - Cellulose

KW - Endoglucanase

KW - Product specificity

KW - Saccharification

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ER -

Effect of a Single Active-Site Cleft Mutation upon Product Specificity in a Thermostable Bacterial Cellulase

Abstract

Bibliographical note

NREL Publication Number

Keywords

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