Comparison of three different promoter systems for secretory α‐amylase production in fed‐batch cultures of recombinant <i>Saccharomyces cerevisiae</i>

Park, Yong Soo; Shiba, Sumihisa; lijima, Shinji; Kobayashi, Takeshi; Hishinuma, Fumio

doi:10.1002/bit.260410904

Cited by 25 publications

(11 citation statements)

References 25 publications

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“…1.6-fold higher than that observed in the batch culture to which only methanol was added. Previously, we reported this level of mouse a-amylase production in a fed-batch culture of recombinant S. cerevisiae 20B-12 [21]. The production rates of a-amylase under the control of the phosphoglycerate kinase (PGK) gene promoter and S. cerevisiae invertase (SUC2) gene promoter were 3.0 and 4.3 kU/l/h, respectively.…”

Section: Discussionmentioning

confidence: 95%

Enhanced production of mouse α-amylase by feeding combined nitrogen and carbon sources in fed-batch culture of recombinant Pichia pastoris

Choi¹,

Park

2006

Process Biochemistry

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The optimal culture conditions for attaining high levels of expression of mouse a-amylase in the methylotrophic yeast Pichia pastoris were investigated. Cell growth and enzyme production were found to be optimal at a temperature of 28 8C and a pH of 6.0. Yeast extract and peptone were the most appropriate nitrogen sources for the expression of a-amylase. When a mixture of 20 g/l peptone and 20 g/l yeast extract was used as the nitrogen source, a-amylase activity reached 300 U/ml. The expression of a-amylase in a fed-batch culture using methanol as the sole carbon source reached 720 U/ml at 3 days of culture. However, when a methanol-to-glycerol mixture was used at ratio of 1:0.5 at a feed rate of 3 g/h, the maximum a-amylase activity was 1.14 kU/ml. Under the optimal culture conditions, when 40 g/l of a mixture of yeast extract and peptone (1:1) was added to the culture at 3 and 5 days the final a-amylase activity increased to 2.4 kU/ml, which was 5.8-and 2.1-fold higher than the activity in the batch culture and in the fed-batch culture with feeding a mixture of methanol and glycerol at a ratio of 1:0.5, respectively. This study demonstrates that an addition of the appropriate amount of an organic nitrogen source to the fed-batch culture is effective to improve high level of expression of mouse a-amylase in the methylotrophic yeast Pichia pastoris. #

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Section: Discussionmentioning

confidence: 95%

Enhanced production of mouse α-amylase by feeding combined nitrogen and carbon sources in fed-batch culture of recombinant Pichia pastoris

Choi¹,

Park

2006

Process Biochemistry

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“…Park et al (1993) compared constitutive promoters P PGK1 and P SUC2 and galactose-inducible promoter P GAL7 for the production of secreted a-amylase in fed-batch cultures. Park et al (1993) compared constitutive promoters P PGK1 and P SUC2 and galactose-inducible promoter P GAL7 for the production of secreted a-amylase in fed-batch cultures.…”

Section: Comparisons Of Promoter Strengthmentioning

confidence: 99%

Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae

2012

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Metabolic pathway engineering in the yeast Saccharomyces cerevisiae leads to improved production of a wide range of compounds, ranging from ethanol (from biomass) to natural products such as sesquiterpenes. The introduction of multienzyme pathways requires precise control over the level and timing of expression of the associated genes. Gene number and promoter strength/regulation are two critical control points, and multiple studies have focused on modulating these in yeast. This MiniReview focuses on methods for introducing genes and controlling their copy number and on the many promoters (both constitutive and inducible) that have been successfully employed. The advantages and disadvantages of the methods will be presented, and applications to pathway engineering will be highlighted.

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“…In Saccharomyces cerevisiae, genes encoding glycolytic enzymes are expressed at a very high level. Thus, promoters of some glycolytic genes have been used to direct the high-level expression of heterologus genes (Park et al 1993).…”

Section: Introductionmentioning

confidence: 99%

Comprehensive cloning and expression analysis of glycolytic genes from the filamentous fungus, Aspergillus oryzae

Nakajima

Kunihiro²,

Sano³

et al. 2000

Current Genetics

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We cloned all the glycolytic genes from Aspergillus oryzae and analyzed their transcriptional regulation by the carbon source in the medium. The deduced amino-acid sequences of the glycolytic genes showed high identity (approximately 41-93%) to those from other lower eukaryotes. Genomic Southern hybridization indicated that all the genes existed as a single copy in the genome. Comparison of mRNA levels between mycelia grown on glucose and on pyruvate showed that most of the A. oryzae glycolytic genes were induced by glucose in the medium. The overall expression profiles of the A. oryzae glycolytic genes resembled those of Saccharomyces cerevisiae. The expression of one of the phosphofructokinase genes (pfkB), however, was repressed by glucose while both PFK1 and PFK2 were induced in S. cerevisiae. These findings indicate that further analysis of the transcriptional regulation of the A. oryzae glycolytic genes will be useful for investigating the evolutionary change of transcription regulation in lower eukaryotes and to construct promoters for industrial applications.

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Comparison of three different promoter systems for secretory α‐amylase production in fed‐batch cultures of recombinant Saccharomyces cerevisiae

Cited by 25 publications

References 25 publications

Enhanced production of mouse α-amylase by feeding combined nitrogen and carbon sources in fed-batch culture of recombinant Pichia pastoris

Enhanced production of mouse α-amylase by feeding combined nitrogen and carbon sources in fed-batch culture of recombinant Pichia pastoris

Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae

Comprehensive cloning and expression analysis of glycolytic genes from the filamentous fungus, Aspergillus oryzae

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