Abstract: Carbohydrate can be used as a substrate that can be consumed by C. butyricum and converted to useful chemicals such as acetic and butyric acid. Influence of concentration and types of carbohydrate to cell growth, carbohydrate consumed, productivity and carbon balance have been explored. Batch reactor was selected in this study to avoid contamination due to simpler operation system. Glucose was preferred as first types of carbohydrate to be tested. Six concentrations were studied from 0 to 28g/L. Eventually, 15g/L has shown the best concentration for glucose in term of growth rate (2.63h-1) and carbon balance (99.76% recovery). Comparison for types of carbohydrate was also conducted. 15g/L of xylose (monosaccharide) and starch (complex carbohydrate) was tested. In term of growth rate and productivity, glucose showed the best carbohydrates. Results for this study showed that glucose and xylose produced more than 80% of acetic acid and less than 20% of butyric acid. Meanwhile, 63.1% of acetic acid and 36.9% of butyric acid were produced from starch.
Abstract: In this study, a synthetic pathway was created by
assembling genes from Clostridium butyricum and Escherichia coli
in different combinations. Among the genes were dhaB1 and dhaB2
from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt)
and its activator (GDHtAc), respectively, involved in the conversion
of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene
from E.coli BL21 was also included which codes for an NADPHdependent
1,3-propanediol oxidoreductase isoenzyme (PDORI)
reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling
analysis indicated that the conformation of fusion protein of YQHD
and DHAB1 was favorable for direct molecular channeling of the
intermediate 3-HPA. According to the simulation results, the yqhD
and dhaB1 gene were assembled in the upstream of dhaB2 to express
a fusion protein, yielding the recombinant strain E. coliBL21
(DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain
BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21
(DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing
the recombinant enzymes simultaneously but in a non-fusion mode.
This is the first report using a gene fusion approach to enhance the
biological conversion of glycerol to the value added compound 1,3-
PD.