PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo

Sang Jun Sim, Kristi D. Snell, Scott A. Hogan, Joanne Stubbe, Chokyun Rha, Anthony J. Sinskey

Research output: Contribution to journalArticle

146 Citations (Scopus)

Abstract

A synthetic operon for polyhydroxyalkanoate (PHA) biosynthesis designed to yield high levels of PHA synthase activity in vivo was constructed by positioning a genetic fragment encoding β-ketothiolase and acetoacetyl-CoA reductase behind a modified synthase gene containing an Escherichia coli promoter and ribosome binding site. Plasmids containing the synthetic operon and the native Alcaligenes eutrophus PHA operon were transformed into E. coli DH5α and analyzed for polyhydroxybutyrate production. The molecular weight of polymer isolated from recombinant E. coli containing the modified synthase construct, determined by multiangle light scattering, was lower than that of the polymer from E. coli containing the native A. eutrophus operon. A further decrease in polyester molecular weight was observed with increased induction of the PHA biosynthetic genes in the synthetic operon. Comparison of the enzyme activity levels of PHA biosynthetic enzymes in a strain encoding the native operon with a strain possessing the synthetic operon indicates that the amount of polyhydroxyalkanoate synthase in a host organism plays a key role in controlling the molecular weight and the polydispersity of polymer.

Original languageEnglish
Pages (from-to)63-67
Number of pages5
JournalNature Biotechnology
Volume15
Issue number1
Publication statusPublished - 1997 Jan 20

Fingerprint

Polyhydroxyalkanoates
Polydispersity
Operon
Escherichia coli
Molecular Weight
Molecular weight
Polymers
Genes
Polyesters
Biosynthesis
Enzyme activity
Binding sites
Light scattering
Plasmids
Enzymes
Alcaligenes
Synthetic Genes
Binding Sites
poly(3-hydroxyalkanoic acid) synthase
Ribosomes

Keywords

  • molecular weight
  • PHA synthase
  • polyhydroxyalkanoates

ASJC Scopus subject areas

  • Microbiology

Cite this

Sim, S. J., Snell, K. D., Hogan, S. A., Stubbe, J., Rha, C., & Sinskey, A. J. (1997). PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo. Nature Biotechnology, 15(1), 63-67.

PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo. / Sim, Sang Jun; Snell, Kristi D.; Hogan, Scott A.; Stubbe, Joanne; Rha, Chokyun; Sinskey, Anthony J.

In: Nature Biotechnology, Vol. 15, No. 1, 20.01.1997, p. 63-67.

Research output: Contribution to journalArticle

Sim, SJ, Snell, KD, Hogan, SA, Stubbe, J, Rha, C & Sinskey, AJ 1997, 'PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo', Nature Biotechnology, vol. 15, no. 1, pp. 63-67.
Sim, Sang Jun ; Snell, Kristi D. ; Hogan, Scott A. ; Stubbe, Joanne ; Rha, Chokyun ; Sinskey, Anthony J. / PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo. In: Nature Biotechnology. 1997 ; Vol. 15, No. 1. pp. 63-67.
@article{a42b648a1cdd4a59ace9557fb7a0f9b5,
title = "PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo",
abstract = "A synthetic operon for polyhydroxyalkanoate (PHA) biosynthesis designed to yield high levels of PHA synthase activity in vivo was constructed by positioning a genetic fragment encoding β-ketothiolase and acetoacetyl-CoA reductase behind a modified synthase gene containing an Escherichia coli promoter and ribosome binding site. Plasmids containing the synthetic operon and the native Alcaligenes eutrophus PHA operon were transformed into E. coli DH5α and analyzed for polyhydroxybutyrate production. The molecular weight of polymer isolated from recombinant E. coli containing the modified synthase construct, determined by multiangle light scattering, was lower than that of the polymer from E. coli containing the native A. eutrophus operon. A further decrease in polyester molecular weight was observed with increased induction of the PHA biosynthetic genes in the synthetic operon. Comparison of the enzyme activity levels of PHA biosynthetic enzymes in a strain encoding the native operon with a strain possessing the synthetic operon indicates that the amount of polyhydroxyalkanoate synthase in a host organism plays a key role in controlling the molecular weight and the polydispersity of polymer.",
keywords = "molecular weight, PHA synthase, polyhydroxyalkanoates",
author = "Sim, {Sang Jun} and Snell, {Kristi D.} and Hogan, {Scott A.} and Joanne Stubbe and Chokyun Rha and Sinskey, {Anthony J.}",
year = "1997",
month = "1",
day = "20",
language = "English",
volume = "15",
pages = "63--67",
journal = "Biotechnology",
issn = "0733-222X",
publisher = "Nature Publishing Group",
number = "1",

}

TY - JOUR

T1 - PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo

AU - Sim, Sang Jun

AU - Snell, Kristi D.

AU - Hogan, Scott A.

AU - Stubbe, Joanne

AU - Rha, Chokyun

AU - Sinskey, Anthony J.

PY - 1997/1/20

Y1 - 1997/1/20

N2 - A synthetic operon for polyhydroxyalkanoate (PHA) biosynthesis designed to yield high levels of PHA synthase activity in vivo was constructed by positioning a genetic fragment encoding β-ketothiolase and acetoacetyl-CoA reductase behind a modified synthase gene containing an Escherichia coli promoter and ribosome binding site. Plasmids containing the synthetic operon and the native Alcaligenes eutrophus PHA operon were transformed into E. coli DH5α and analyzed for polyhydroxybutyrate production. The molecular weight of polymer isolated from recombinant E. coli containing the modified synthase construct, determined by multiangle light scattering, was lower than that of the polymer from E. coli containing the native A. eutrophus operon. A further decrease in polyester molecular weight was observed with increased induction of the PHA biosynthetic genes in the synthetic operon. Comparison of the enzyme activity levels of PHA biosynthetic enzymes in a strain encoding the native operon with a strain possessing the synthetic operon indicates that the amount of polyhydroxyalkanoate synthase in a host organism plays a key role in controlling the molecular weight and the polydispersity of polymer.

AB - A synthetic operon for polyhydroxyalkanoate (PHA) biosynthesis designed to yield high levels of PHA synthase activity in vivo was constructed by positioning a genetic fragment encoding β-ketothiolase and acetoacetyl-CoA reductase behind a modified synthase gene containing an Escherichia coli promoter and ribosome binding site. Plasmids containing the synthetic operon and the native Alcaligenes eutrophus PHA operon were transformed into E. coli DH5α and analyzed for polyhydroxybutyrate production. The molecular weight of polymer isolated from recombinant E. coli containing the modified synthase construct, determined by multiangle light scattering, was lower than that of the polymer from E. coli containing the native A. eutrophus operon. A further decrease in polyester molecular weight was observed with increased induction of the PHA biosynthetic genes in the synthetic operon. Comparison of the enzyme activity levels of PHA biosynthetic enzymes in a strain encoding the native operon with a strain possessing the synthetic operon indicates that the amount of polyhydroxyalkanoate synthase in a host organism plays a key role in controlling the molecular weight and the polydispersity of polymer.

KW - molecular weight

KW - PHA synthase

KW - polyhydroxyalkanoates

UR - http://www.scopus.com/inward/record.url?scp=0002217020&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0002217020&partnerID=8YFLogxK

M3 - Article

C2 - 9035108

AN - SCOPUS:0002217020

VL - 15

SP - 63

EP - 67

JO - Biotechnology

JF - Biotechnology

SN - 0733-222X

IS - 1

ER -