Expression and secretion of rice α-amylase by Saccharomyces cerevisiae

Monto H. Kumagai, Mena Shah, Masaaki Terashima, Zeljko Vrkljan, John R. Whitaker, Raymond L. Rodriguez

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

We report the high level expression and secretion of rice α-amylase isozyme by Saccharomyces cerevisiae. Transcription of this gene was under control of the yeast enolase promoter. The synthesized protein had an approximate molecular size of 45 kDa and a pI of approx 4.7 to 5.0. The rice α-amylase signal peptide was recognized and efficiently processed by yeast and the active, glycosylated enzyme was secreted into the culture media. This enzyme was purified to homogeneity by affinity chromotography and its enzymatic properties were characterized. The Km and Vmax were found to be similar to those of α-amylase from other organisms. The high level of secretion observed in these studies may be due to the unique features of the rice signal peptide and/or to the glycosylation of the recombinant enzyme.

Original languageEnglish (US)
Pages (from-to)209-216
Number of pages8
JournalGene
Volume94
Issue number2
DOIs
StatePublished - 1990

Fingerprint

Amylases
Saccharomyces cerevisiae
Protein Sorting Signals
Enzymes
Yeasts
Phosphopyruvate Hydratase
Glycosylation
Isoenzymes
Culture Media
Genes
Oryza
Proteins

Keywords

  • 2 μ plasmid
  • affinity chromatography
  • amylolytic yeast
  • enolase promoter
  • Recombinant DNA
  • signal peptide

ASJC Scopus subject areas

  • Genetics

Cite this

Kumagai, M. H., Shah, M., Terashima, M., Vrkljan, Z., Whitaker, J. R., & Rodriguez, R. L. (1990). Expression and secretion of rice α-amylase by Saccharomyces cerevisiae. Gene, 94(2), 209-216. https://doi.org/10.1016/0378-1119(90)90389-9

Expression and secretion of rice α-amylase by Saccharomyces cerevisiae. / Kumagai, Monto H.; Shah, Mena; Terashima, Masaaki; Vrkljan, Zeljko; Whitaker, John R.; Rodriguez, Raymond L.

In: Gene, Vol. 94, No. 2, 1990, p. 209-216.

Research output: Contribution to journalArticle

Kumagai, MH, Shah, M, Terashima, M, Vrkljan, Z, Whitaker, JR & Rodriguez, RL 1990, 'Expression and secretion of rice α-amylase by Saccharomyces cerevisiae', Gene, vol. 94, no. 2, pp. 209-216. https://doi.org/10.1016/0378-1119(90)90389-9
Kumagai MH, Shah M, Terashima M, Vrkljan Z, Whitaker JR, Rodriguez RL. Expression and secretion of rice α-amylase by Saccharomyces cerevisiae. Gene. 1990;94(2):209-216. https://doi.org/10.1016/0378-1119(90)90389-9
Kumagai, Monto H. ; Shah, Mena ; Terashima, Masaaki ; Vrkljan, Zeljko ; Whitaker, John R. ; Rodriguez, Raymond L. / Expression and secretion of rice α-amylase by Saccharomyces cerevisiae. In: Gene. 1990 ; Vol. 94, No. 2. pp. 209-216.
@article{5bb1d1bb76fd4bc8935dbd7ba469045e,
title = "Expression and secretion of rice α-amylase by Saccharomyces cerevisiae",
abstract = "We report the high level expression and secretion of rice α-amylase isozyme by Saccharomyces cerevisiae. Transcription of this gene was under control of the yeast enolase promoter. The synthesized protein had an approximate molecular size of 45 kDa and a pI of approx 4.7 to 5.0. The rice α-amylase signal peptide was recognized and efficiently processed by yeast and the active, glycosylated enzyme was secreted into the culture media. This enzyme was purified to homogeneity by affinity chromotography and its enzymatic properties were characterized. The Km and Vmax were found to be similar to those of α-amylase from other organisms. The high level of secretion observed in these studies may be due to the unique features of the rice signal peptide and/or to the glycosylation of the recombinant enzyme.",
keywords = "2 μ plasmid, affinity chromatography, amylolytic yeast, enolase promoter, Recombinant DNA, signal peptide",
author = "Kumagai, {Monto H.} and Mena Shah and Masaaki Terashima and Zeljko Vrkljan and Whitaker, {John R.} and Rodriguez, {Raymond L.}",
year = "1990",
doi = "10.1016/0378-1119(90)90389-9",
language = "English (US)",
volume = "94",
pages = "209--216",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Expression and secretion of rice α-amylase by Saccharomyces cerevisiae

AU - Kumagai, Monto H.

AU - Shah, Mena

AU - Terashima, Masaaki

AU - Vrkljan, Zeljko

AU - Whitaker, John R.

AU - Rodriguez, Raymond L.

PY - 1990

Y1 - 1990

N2 - We report the high level expression and secretion of rice α-amylase isozyme by Saccharomyces cerevisiae. Transcription of this gene was under control of the yeast enolase promoter. The synthesized protein had an approximate molecular size of 45 kDa and a pI of approx 4.7 to 5.0. The rice α-amylase signal peptide was recognized and efficiently processed by yeast and the active, glycosylated enzyme was secreted into the culture media. This enzyme was purified to homogeneity by affinity chromotography and its enzymatic properties were characterized. The Km and Vmax were found to be similar to those of α-amylase from other organisms. The high level of secretion observed in these studies may be due to the unique features of the rice signal peptide and/or to the glycosylation of the recombinant enzyme.

AB - We report the high level expression and secretion of rice α-amylase isozyme by Saccharomyces cerevisiae. Transcription of this gene was under control of the yeast enolase promoter. The synthesized protein had an approximate molecular size of 45 kDa and a pI of approx 4.7 to 5.0. The rice α-amylase signal peptide was recognized and efficiently processed by yeast and the active, glycosylated enzyme was secreted into the culture media. This enzyme was purified to homogeneity by affinity chromotography and its enzymatic properties were characterized. The Km and Vmax were found to be similar to those of α-amylase from other organisms. The high level of secretion observed in these studies may be due to the unique features of the rice signal peptide and/or to the glycosylation of the recombinant enzyme.

KW - 2 μ plasmid

KW - affinity chromatography

KW - amylolytic yeast

KW - enolase promoter

KW - Recombinant DNA

KW - signal peptide

UR - http://www.scopus.com/inward/record.url?scp=0025221190&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025221190&partnerID=8YFLogxK

U2 - 10.1016/0378-1119(90)90389-9

DO - 10.1016/0378-1119(90)90389-9

M3 - Article

C2 - 2258052

AN - SCOPUS:0025221190

VL - 94

SP - 209

EP - 216

JO - Gene

JF - Gene

SN - 0378-1119

IS - 2

ER -