• Korean Chemical Engineering Research
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• v.46 no.5
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• pp.858-862
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• 2008

The Simultaneous Saccharification and Fermentation(SSF) of ethanol production from potato starch studied with respect to growth pH, temperature, substrate concentration. The glucoamylase and Saccharomyceses cerevisiae have a capacity to carry out a single stage SSF process for ethanol production. The characteristics, termed as starch hydrolysis, accumulation of glucose, ethanol production and biomass formation, were affected with variation in pH, temperature and starch concentration. The maximum ethanol concentration of 12.9g/l was obtained using a starch concentration 30g/l, which represent an ethanol yield of 86%. The optimum conditions for the maximum ethanol yield were found to be a temperature of 38, pH of 4.0 and fermentation time of 18hr. Thus by using the control composite design, it is possible to determine the accurate values of the fermentation parameters where maximum production of ethanol occurs.

• BMB Reports
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• v.37 no.4
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• pp.429-438
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• 2004

Complementary DNAs encoding $\alpha$-amylases (Amyl I, Amyl III) and glucoamylase (GA I) were cloned from Aspergillus awamori KT-11 and their nucleotide sequences were determined. The sequence of Amyl III that was a raw starch digesting $\alpha$-amylase was found to consist of a 1,902 bp open reading frame encoding 634 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. On the other hand, the sequence of Amyl I, which cannot act on raw starch, consisted of a 1,500 bp ORF encoding 499 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. The sequence of GA I consisted of a 1,920 bp ORF that encoded 639 amino acids. The signal peptide was composed of 24 amino acids. The amino acid sequence of Amyl III from the N-terminus to the amino acid number 499 showed 63.3% homology with Amyl I. However, the amino acid sequence from the amino acid number 501 to C-terminus, including the raw-starch-affinity site and the TS region rich in threonine and serine, showed 66.9% homology with GA I.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1401-1405
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• 2012

A thermotolerant Saccharomyces cerevisiae mutant strain, TT6, was constructed after multi-parental hybridization of five mutant strains obtained by UV or NTG treatment of the original strain, S. cerevisiae KV1. When incubated at $40^{\circ}C$ in YPD broth, TT6 began to grow exponentially in 10 h, but KV1 did not show any noticeable growth even after 22 h. The thermotolerant growth of TT6 was confirmed by serial dilution assay at $42^{\circ}C$; TT6 grew at a cell concentration ($10^{-5}$) 10,000 times lower than that of KV1 ($10^{-1}$). Whereas ethanol production from YP containing 23% (w/v) glucose by KV1 decreased with increasing temperature from $30^{\circ}C$ to $36^{\circ}C$, ethanol production by TT6 did not decrease at temperatures up to $37^{\circ}C$. When TT6 was tested for ethanol production at $36^{\circ}C$ by simultaneous saccharification and fermentation (SSF) from 23% corn, 24 h of fermentation time or 50% of the glucoamylase dose was saved when compared with KV1 at $30^{\circ}C$. The ethanol yield from corn by SSF with TT6 at $36^{\circ}C$ was 91.7% of the theoretical yield, whereas that of KV1 at $30^{\circ}C$ was 90.6%.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.120-127
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• 1990

Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.166-172
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• 1997

Aspergillus niger was selected as a strain producing the potent raw starch hydorlyzing enzyme. These experiments were conducted to investigate the conditions of the glucoa- mylase production, the purification of the enzyme, some characteristics of the purified enzyme and hydrolysis rate on various raw starches such as com, rice, potato, glutinous rice, sweet potato, wheat and barley. The optimum cultural temperature and time for the enzyme production on wheat bran medium were $30^{\circ}C$ and 96hrs, respectively. The respective addition of yeast extract and nutrient broth on wheat bran medium increased slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 30.7u/mg-protein and the yield of enzyme activity was 25.8%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 56,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pH3.7. The optimum temperature and pH were $65^{\circ}C$ and pH 4.0, respectively. The purified enzyme was stable in the pH range of pH 3.0-9.5 and below $45^{\circ}C$, and its thermal stability was slightly increased by the addition of $Ca^{2+}$. The purified enzyme was activated by $Co^{2+},\;Sr^{2+},\;Mn^{2+},\;Fe^{2+},\;Cu^{2+}$. Raw rice starch, raw corn starch, raw glutinous rice starch, raw sweet potato starch, raw wheat starch and raw barley starch showed more than 90% hydrolysis rate in 48hrs incubation. Even raw potato starch, most difficult to be hydrolyzed, showed 80% hydrolysis rate. The purified enzyme was identified as glucoamylase.

• Microbiology and Biotechnology Letters
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• v.3 no.3
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• pp.147-156
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• 1975

The possibility of using tapioca pellets as a raw material in glutanmic acid fermentation by Microcuccus glutamicus is shown. The ground pellets were diluted with water to 20％ solid level and treated with $\alpha$-anylase prepared from a thermophilic Actinomycetes strain culture for 90 min at 85$^{\circ}C$ under pH 6.0. The liquefied solution was further saccharified with commercial glucoamylase for 36 hours under the reaction conditions of 55$^{\circ}C$ and pH 5.0. The inhibitory effect of excess biotin content, 16 $\mu\textrm{g}$ Per liter of the hydrolzate, could be reduced effectively by adding 10 IU of penicillin per ml of the medium after five hours of the fermentation. The maximum glutamic acid yield of 38.5 g/l was obtained after 60 hours of shaking culture at 28-3$0^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.415-420
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• 1986

Alcohol fermentation of uncooked naked barley was carried out by the combined action of the maceration enzyme from black Aspergillus niger and the glucoamylase from Rhizopus sp. The combined enzyme preparation was found to be effective in maceration and saccharification of the raw naked barley starch. The Hydrolysis rate measured by the amount of glucose liberated reached more than 70% at pH 4.5 and 3$0^{\circ}C$ after 76 hrs. For alcohol fermentation without cooking, the naked barley mash of 18% initial total sugar was pretreated with concentrated sulfuric acid (0.15 weight % of the mash volume) at 55$^{\circ}C$ for 2hr, and used for alcohol fermentation. A simultaneous saccharification and fermentation was carried out at pH 4.8 and 3$0^{\circ}C$ for 96 hrs. Under this fermentation condition, 3.5% increase in alcohol yield together with 2.0% increase in alcohol concentration were obtained when compared with the conventional cooking fermentation.

• Korean Journal of Food Science and Technology
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• v.22 no.3
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• pp.296-299
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• 1990

As an improved preparation method of Shikhae, a tea-bag system containing malt and amylolytic enzymes was developed in which extraction of malt enzymes and saccharification occured efficiently. The amylolytic activity of the malt was increased by adding the mixture of ${\alpha}-amylase$, glucoamylase and glucoisomerase. Malt and the mixture of enzymes were placed in tea-bag $(16{\times}20cm)$, extracted in water at $30-40^{\circ}C$ for 1-2 hours and followed by saccharification of the cooked rice at $60-70^{\circ}C$ for 3-4 hours. In the conventional Shikhae, content of maltose was about 50% and that of oligosaccharides larger than trisaccharides was about 40% of total sugar. The content of monosaccharides such as glucose and fructose was about 95% and this improved method would be effective for increasing the sweetness and the monosaccharide contents in the product.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.427-429
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• 1996

We have already cloned an extracellular $\alpha$-glucosidase gene from Aspergillus niger with oligonucleotide probe synthesized on the basis of the peptide sequences determined previously. The DNA sequence revealed an open reading frame of 895 amino acids split by three introns. We are attempting to construct an A. niger strain deficient in the $\alpha$-glucosidase enzyme activity, which would be useful for the glucoamylase production without contamination by the industrially undesirable $\alpha$-glucosidase. For destruction of the $\alpha$-glucosidase gene, we try to make transformations. A cloned partial $\alpha$-glucosidase gene was introduced into Aspergillus niger, and transformants with suppressed $\alpha$-glucosidase activity were isolated. The transformants were cultured on YPD medium which contained Hygromycin B at 30$\circ$C. The activity of $\alpha$-glucosidase of the suppressed transformants was compared to that of wild type activity. As shown by southern-hybridization, we detected that the transformant was a heterocaryon.

• Journal of the Korean Professional Engineers Association
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• v.17 no.1
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• pp.27-35
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• 1984

When starch is hydrolyzed either by acid or by the enzymes maltase or diastase, contained in germinating barley, a yield of 80% of maltose is obtained. Maltose is built of two molecules of ${\alpha}$-glucose, bound in the position 1:4 i.e., carbon atom 1 of one glucose molecule is bound in a glucosidic bond to carbon atom 4 of the second molecule. Until around 1960, dextrose and glucose syrups were prepared from starch exclusively by acid hydrolysis. The process was corrosive, and the dextrose yield low. It was, therefore, a great step forward when pure glucoamylase in combination with bacterial ${\alpha}$-amylase made possible a complete enzymatic hydrolysis of starch to dextrose. Today several enzymatic processes are used in the industry.