Initiation, Elongation, and Termination of Bacterial Cellulose Synthesis

John B. McManus, Hui Yang, Liza Anne Wilson, James D. Kubicki, Ming Tien

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Cellulose is the major component of the plant cell wall and composed of β-linked glucose units. Use of cellulose is greatly impacted by its physical properties, which are dominated by the number of individual cellulose strand within each fiber and the average length of each strand. Our work described herein provides a complete mechanism for cellulose synthase accounting for its processivity and mechanism of initiation. Using ionic liquids and gel permeation chromatography, we obtain kinetic constants for initiation, elongation, and termination (release of the cellulose strand from the enzyme) for two bacterial cellulose synthases (Gluconacetobacter hansenii and Rhodobacter sphaeroides). Our results show that initiation of synthesis is primer-independent. After initiation, the enzyme undergoes multiple cycles of elongation until the strand is released. The rate of elongation is much faster than that of steady-state turnover. Elongation requires cyclic addition of glucose (from uridine diphosphate-glucose) and then strand translocation by one glucose unit. Translocations greater than one glucose unit result in termination requiring reinitiation. The rate of the strand release, relative to the rate of elongation, determines the processivity of the enzyme. This mechanism and the measured rate constants were supported by kinetic simulation. With the experimentally determined rate constants, we are able to simulate steady-state kinetics and mimic the size distribution of the product. Thus, our results provide for the first time a mechanism for cellulose synthase that accounts for initiation, elongation, and termination.

Original languageEnglish (US)
Pages (from-to)2690-2698
Number of pages9
JournalACS Omega
Volume3
Issue number3
DOIs
StatePublished - Jan 1 2018

Fingerprint

Cellulose
Elongation
Glucose
Enzymes
Kinetics
Rate constants
Uridine Diphosphate Glucose
Ionic Liquids
Gel permeation chromatography
Ionic liquids
Physical properties
Fibers
cellulose synthase

All Science Journal Classification (ASJC) codes

  • Chemical Engineering(all)
  • Chemistry(all)

Cite this

McManus, John B. ; Yang, Hui ; Wilson, Liza Anne ; Kubicki, James D. ; Tien, Ming. / Initiation, Elongation, and Termination of Bacterial Cellulose Synthesis. In: ACS Omega. 2018 ; Vol. 3, No. 3. pp. 2690-2698.
@article{51ca91ef265e4994834d349b39fdf031,
title = "Initiation, Elongation, and Termination of Bacterial Cellulose Synthesis",
abstract = "Cellulose is the major component of the plant cell wall and composed of β-linked glucose units. Use of cellulose is greatly impacted by its physical properties, which are dominated by the number of individual cellulose strand within each fiber and the average length of each strand. Our work described herein provides a complete mechanism for cellulose synthase accounting for its processivity and mechanism of initiation. Using ionic liquids and gel permeation chromatography, we obtain kinetic constants for initiation, elongation, and termination (release of the cellulose strand from the enzyme) for two bacterial cellulose synthases (Gluconacetobacter hansenii and Rhodobacter sphaeroides). Our results show that initiation of synthesis is primer-independent. After initiation, the enzyme undergoes multiple cycles of elongation until the strand is released. The rate of elongation is much faster than that of steady-state turnover. Elongation requires cyclic addition of glucose (from uridine diphosphate-glucose) and then strand translocation by one glucose unit. Translocations greater than one glucose unit result in termination requiring reinitiation. The rate of the strand release, relative to the rate of elongation, determines the processivity of the enzyme. This mechanism and the measured rate constants were supported by kinetic simulation. With the experimentally determined rate constants, we are able to simulate steady-state kinetics and mimic the size distribution of the product. Thus, our results provide for the first time a mechanism for cellulose synthase that accounts for initiation, elongation, and termination.",
author = "McManus, {John B.} and Hui Yang and Wilson, {Liza Anne} and Kubicki, {James D.} and Ming Tien",
year = "2018",
month = "1",
day = "1",
doi = "10.1021/acsomega.7b01808",
language = "English (US)",
volume = "3",
pages = "2690--2698",
journal = "ACS Omega",
issn = "2470-1343",
publisher = "American Chemical Society",
number = "3",

}

Initiation, Elongation, and Termination of Bacterial Cellulose Synthesis. / McManus, John B.; Yang, Hui; Wilson, Liza Anne; Kubicki, James D.; Tien, Ming.

In: ACS Omega, Vol. 3, No. 3, 01.01.2018, p. 2690-2698.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Initiation, Elongation, and Termination of Bacterial Cellulose Synthesis

AU - McManus, John B.

AU - Yang, Hui

AU - Wilson, Liza Anne

AU - Kubicki, James D.

AU - Tien, Ming

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Cellulose is the major component of the plant cell wall and composed of β-linked glucose units. Use of cellulose is greatly impacted by its physical properties, which are dominated by the number of individual cellulose strand within each fiber and the average length of each strand. Our work described herein provides a complete mechanism for cellulose synthase accounting for its processivity and mechanism of initiation. Using ionic liquids and gel permeation chromatography, we obtain kinetic constants for initiation, elongation, and termination (release of the cellulose strand from the enzyme) for two bacterial cellulose synthases (Gluconacetobacter hansenii and Rhodobacter sphaeroides). Our results show that initiation of synthesis is primer-independent. After initiation, the enzyme undergoes multiple cycles of elongation until the strand is released. The rate of elongation is much faster than that of steady-state turnover. Elongation requires cyclic addition of glucose (from uridine diphosphate-glucose) and then strand translocation by one glucose unit. Translocations greater than one glucose unit result in termination requiring reinitiation. The rate of the strand release, relative to the rate of elongation, determines the processivity of the enzyme. This mechanism and the measured rate constants were supported by kinetic simulation. With the experimentally determined rate constants, we are able to simulate steady-state kinetics and mimic the size distribution of the product. Thus, our results provide for the first time a mechanism for cellulose synthase that accounts for initiation, elongation, and termination.

AB - Cellulose is the major component of the plant cell wall and composed of β-linked glucose units. Use of cellulose is greatly impacted by its physical properties, which are dominated by the number of individual cellulose strand within each fiber and the average length of each strand. Our work described herein provides a complete mechanism for cellulose synthase accounting for its processivity and mechanism of initiation. Using ionic liquids and gel permeation chromatography, we obtain kinetic constants for initiation, elongation, and termination (release of the cellulose strand from the enzyme) for two bacterial cellulose synthases (Gluconacetobacter hansenii and Rhodobacter sphaeroides). Our results show that initiation of synthesis is primer-independent. After initiation, the enzyme undergoes multiple cycles of elongation until the strand is released. The rate of elongation is much faster than that of steady-state turnover. Elongation requires cyclic addition of glucose (from uridine diphosphate-glucose) and then strand translocation by one glucose unit. Translocations greater than one glucose unit result in termination requiring reinitiation. The rate of the strand release, relative to the rate of elongation, determines the processivity of the enzyme. This mechanism and the measured rate constants were supported by kinetic simulation. With the experimentally determined rate constants, we are able to simulate steady-state kinetics and mimic the size distribution of the product. Thus, our results provide for the first time a mechanism for cellulose synthase that accounts for initiation, elongation, and termination.

UR - http://www.scopus.com/inward/record.url?scp=85043316615&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85043316615&partnerID=8YFLogxK

U2 - 10.1021/acsomega.7b01808

DO - 10.1021/acsomega.7b01808

M3 - Article

VL - 3

SP - 2690

EP - 2698

JO - ACS Omega

JF - ACS Omega

SN - 2470-1343

IS - 3

ER -