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Production of
1,3-propanediol from glycerol by engineered Escherichia
coli using a novel co-expression vector
Zheng Ma1,
Zhiming Rao1*, Liyu Xu2, Xiangru Liao1, Huiying Fang1,
Bin Zhuge1 and Jian Zhuge1
1The
Key Laboratory of Industrial biotechnology of Ministry of
Education, Research Center of Industrial Microbiology,
School of Biotechnology, Jiangnan University, Wuxi, Jiangsu
Province 214122, P.R. China.
2Applied
Science Department, Zhejiang Economic and Trade Polytechnic,
Hangzhou 310018, China.
*Corresponding author.
E-mail:
raozm@yahoo.com.cn.
Tel:
+86-510-85918109.
Fax: +86-510-85918106.
Accepted
11 September, 2009 |
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1,3-Propanediol (1,3-PD) has versatile applications in
polymers, cosmetics, foods and medicines. In order to
consolidate the functions of glycerol dehydratase gene
dhaB and 1,3-propanediol oxidoreductase gene dhaT
and produce
1,3-PD from glycerol, the genes dhaB and dhaT
from Klebsiella pneumoniae were inserted into a
co-expression vector pACYCDuet-1 synchronously and the
recombinant strain E. coli/pACYCDuet-dhaB-dhaT
was obtained.
Both
enzymes were functionally co-expressed in E. coli at
the presence of the selective pressure and the addition of
the IPTG. The specific enzyme activity of DHAB and DHAT were
8.3 and 6.2 U/mg, respectively. When cultivated at 37°C for
30 h, the recombinant microorganisms produced 1,3-PD of 11.3
g with the consumption of 40 g glycerol per liter. The
production of 1,3-PD by the strain
E. coli/pACYCDuet-dhaB-dhaT
was about 13-fold higher than the recombinant
E. coli
harboring the gene dhaB.
Key
words:
1,3-propanediol, Klebsiella pneumoniae, co-expression
system, T7 RNA polymerase promoter. |
