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Metabolic engineering of Rhizopus oryzae: Effects of overexpressing pyc and pepc genes on fumaric acid biosynthesis from glucose

Paper ID Volume ID Publish Year Pages File Format Full-Text
31565 44819 2012 9 PDF Available
Title
Metabolic engineering of Rhizopus oryzae: Effects of overexpressing pyc and pepc genes on fumaric acid biosynthesis from glucose
Abstract

Fumaric acid, a dicarboxylic acid used as a food acidulant and in manufacturing synthetic resins, can be produced from glucose in fermentation by Rhizopus oryzae. However, the fumaric acid yield is limited by the co-production of ethanol and other byproducts. To increase fumaric acid production, overexpressing endogenous pyruvate carboxylase (PYC) and exogenous phosphoenolpyruvate carboxylase (PEPC) to increase the carbon flux toward oxaloacetate were investigated. Compared to the wild type, the PYC activity in the pyc transformants increased 56%–83%, whereas pepc transformants exhibited significant PEPC activity (3–6 mU/mg) that was absent in the wild type. Fumaric acid production by the pepc transformant increased 26% (0.78 g/g glucose vs. 0.62 g/g for the wild type). However, the pyc transformants grew poorly and had low fumaric acid yields (<0.05 g/g glucose) due to the formation of large cell pellets that limited oxygen supply and resulted in the accumulation of ethanol with a high yield of 0.13–0.36 g/g glucose. This study is the first attempt to use metabolic engineering to modify the fumaric acid biosynthesis pathway to increase fumaric acid production in R. oryzae.

► Fumaric acid synthesis pathway in R. oryzae was engineered to increase flux toward oxaloacetate. ► Pyruvate carboxylase (PYC) and phosphoenolpyruvate carboxylase (PEPC) were overexpressed. ► Overexpressing PYC resulted in poor growth and fumaric acid production due to reduced ATP. ► Overexpressing PEPC increased fumaric acid production from glucose by 26%. ► This is the first metabolic engineering study to increase fumaric acid production in R. oryzae.

Keywords
Fumaric acid; Phosphoenolpyruvate carboxylase; Pyruvate carboxylase; Metabolic engineering; Rhizopus oryzae
First Page Preview
Metabolic engineering of Rhizopus oryzae: Effects of overexpressing pyc and pepc genes on fumaric acid biosynthesis from glucose
Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 14, Issue 5, September 2012, Pages 512–520
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering