• Journal of Microbiology and Biotechnology
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• 제3권2호
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• pp.99-107
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• 1993

Two strains (No. 26 and 143) of bacteria which secrete both pectinase and raw-starch-digesting amylase simultaneously, were isolated from various domestic soil samples. The two bacteria were identified as Pasteurella ureae judging by their morphological and physiological characteristics. The optimal culture conditions for the production of raw-starch-digesting enzyme by the Pasteurella ureae 26 were using $NH_4NO_3$ as the nitrogen source at $37^{\circ}C$ with the pH of 7.5, and 15 of C/N ratio. Since the enzyme was produced only when raw or soluble starch was used as a carbon source, but not when glucose or other sugars was used, the enzyme was considered to be an inducible enzyme by starch. Thin layer chromatography of the hydrolyzed product of starch by the raw-starch-digesting enzyme of the strain No. 26 showed that glucose, maltose and other oligosaccharides were present in the hydrolyzates, and therefore the enzyme seemed to be ${\alpha}-amylase$. The enzyme had adsorbability onto raw com starch in the pH range of 3 to 9.

• Korean Journal of Food Science and Technology
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• 제27권5호
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• pp.773-775
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• 1995

The effect of alum$(Al{\cdot}K(SO_4)_2{\cdot}12H_{2}O)$ on the activity of raw starch-digesting enzyme produced by alkali-tolerant Bacillus sp. was investigated. In adding alum of 0.5%(w/w), activities of raw starch-digesting enzyme on the gelatinized and raw rice starches have not been inhibited. In case of adding alum of 5%(w/w), competitive and uncompetitive inhibition were observed for the gelatinized and raw rice starches, respectively. The inhibitory effect on the raw starch was much higher than that on the gelatinized starch.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.718-726
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• 2010

A selected fungal strain, for production of the raw-starchdigesting enzyme by solid-state fermentation, was improved by two repeated sequential exposures to ${\gamma}$-irradiation of $Co^{60}$, ultraviolet, and four repeated treatments with Nmethyl-N'-nitrosoguanidine. The mutant strain Aspergillus sp. XN15 was chosen after a rigorous screening process, with its production of the raw-starch-digesting enzyme being twice that of usual wild varieties cultured under preoptimized conditions and in an unsupplemented medium. After 17 successive subculturings, the enzyme production of the mutant was stable. Optimal conditions for the production of the enzyme by solid-state fermentation, using wheat bran as the substrate, were accomplished for the mutant Aspergillus sp. XN15. With the optimal fermentation conditions, and a solid medium supplemented with nitrogen sources of 1% urea and 1% $NH_4NO_3$, 2.5 mM $CoSO_4$, 0.05% (v/w) Tween 80, and 1% glucose, the mutant Aspergillus sp. XN15 produced the raw-starch-digesting enzyme in quantities 19.4 times greater than a typical wild variety. Finally, XN15, through simultaneous saccharification and fermentation of a raw rice corn starch slurry, produced a high level of ethanol with $Y_{p/s}$ of 0.47 g/g.

• KSBB Journal
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• 제13권5호
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• pp.621-625
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• 1998

The raw starch hydrolysis by amylase prepared from alkaline-tolerant Bacillus sp. were investigated. Degree of hydrolysis(%) of 5%(w/v) raw rice, corn and potato starch by this enzyme were about 40, 25 and 20%, respectively. The hydrolysis action on raw starch by change of blue value was similar to the action pattern of exo ${\beta}$-amylase. The hydrolysis products of rice starch were mainly glucose and maltose. Oligosaccarides were also detected. From the above results, this enzyme was considered as exo type ${\alpha}$-amylase. This enzyme activity on the raw starch and the gelatinized starch were 28.40 and 86.60 IU/mg protein, respectively, and the ratio of raw starch-digesting activity to gelatinized starch-digesting activity (raw starch digestivity) was about 32%. The Km values for the raw and the gelatinized starch were 4.22 and 3.0mg/mL, respectively, and the VmaX values were 0.20 and 0.31mg/mL/min, respectively.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1161-1168
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• 2009

Ethanol production by the simultaneous saccharification and fermentation (SSF) of low-value rice wine cake (RWC) without cooking was investigated. RWC is the filtered solid waste of fermented rice wine mash and contains 53% raw starch. For the SSF, the RWC slurry was mixed with the raw-starch-digesting enzyme of Rhizopus sp. and yeast, where the yeast strain was selected from 300 strains and identified as Saccharomyces cerevisiae KV25. The highest efficiency (94%) of ethanol production was achieved when the uncooked RWC slurry contained 23.03% starch. The optimal SSF conditions were determined as 1.125 units of the raw-starch-digesting enzyme per gram of RWC, a fermentation temperature of $30^{\circ}C$, slurry pH of 4.5, 36-h-old seeding culture, initial yeast cell number of $2{\times}10^7$ per ml of slurry, 17 mM of urea as the nitrogen additive, 0.25 mM of $Cu^{2+}$ as the metal ion additive, and a fermentation time of 90 h. Under these optimal conditions, the ethanol production resulting from the SSF of the uncooked RWC slurry was improved to 16.8% (v/v) from 15.1% (v/v) of pre-optimization.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.383-390
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• 2010

A newly isolated active producer of raw-starch-digesting amyloytic enzymes, Rhizopus microsporus var. chinensis CICIM-CU F0088, was screened and identified by morphological characteristics and molecular phylogenetic analyses. This fungus was isolated from the soil of Chinese glue pudding mill, and produced high levels of amylolytic activity under solid-state fermentation with supplementation of starch and wheat bran. Results of thin-layer chromatography showed there are two kinds of amyloytic enzymes formed by this strain, including one $\alpha$-amylase and two glucoamylases. It was found in the electron microscope experiments that the two glucoamylases can digest raw corn starch and have an optimal temperature of $70^{\circ}C$. These results signified that amyloytic enzymes secreted by strain Rhizopus microsporus var. chinensis CICIM-CU F0088 were types of thermostable amyloytic enzymes and able to digest raw corn starch.

• Journal of the korean academy of Pediatric Dentistry
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• 제23권3호
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• pp.764-773
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• 1996

The mutan containing $\alpha$-1,3 bond is an insoluble portion of glucan which is the main component of dental plaque. The secretion of mutanase was assessed with mutan-digesting Streptomyces isolated from soil, and the factors affecting its activity was studied, obtaining the following result. Mutan-digesting Streptomyces was identified as Streptomyces exfoliatus by its characteristics. The effect of dextranase was identified on the media containing blue dextran. A clear zone was produced by Streptomyces exfoliatus on the media containing blue mutan, so showing the secretion of mutanase. A clear zone was significantly produced on the media overlayed with agar containing blue mutan. A clear zone was produced at 2 days after the inoculation of Streptomyces exfoliatus on the media containing below a concentration of 0.025% glucose, at 3 days on the media containing 0.05 % glucose, and at 4 days on the media containing 0.1 % glucose. Mutan-digestion wasn't appeared early by adding other carbohydrates. The higher concentration of peptone, the later appearance of clear zone was on the media containing below a concentration of 0.1 % peptone. These results indicated that the secretion of mutanase was identified from mutan-digesting Streptomyces on the media containing blue mutan, and a clear zone was appeared lately on the media containing higher amount of glucose.

• Journal of Microbiology and Biotechnology
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• 제2권1호
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• pp.56-65
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• 1992

The complete nucleotide sequence of the Bacillus circulans F-2 RSDA gene, coding for raw starch digesting a-amylase (RSDA), has been determined. The RSDA structure gene consists of an open reading frame of 2508 bp. Six bp upstream of the translational start codon of the RSDA is a typical gram-positive Shine-Dalgarno sequence and the RSDA encodes a preprotein of 836 amino acids with an Mr of 96, 727. The gene was expressed from its own regulatory region in E. coli and two putative consensus promoter sequences were identified upstream of a ribosome binding site and an ATG start codon. Confirmation of the nucleotide sequence was obtained and the signal peptide cleavage site was identified by comparing the predicted amino acid sequence with that derived by N-terminal analysis of the purified RSDA. The deduced N-terminal region of the RSDA conforms to the general pattern for the signal peptides of secreted prokaryotic proteins. The complete amino acid sequence was deduced and homology with other enzymes was compared. The results suggested that the Thr-Ser-rich hinge region and the non-catalytic domain are necessary for efficient adsorption onto raw substrates, and the catalytic domain (60 kDa) is necessary for the hydrolysis of substrates, as suggested in previous studies (8, 9).

• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1542-1550
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• 2014

Co-digesting molasses wastewater and sewage sludge was evaluated for hydrogen production by response surface methodology (RSM). Batch experiments in accordance with various dilution ratios (40- to 5-fold) and waste mixing composition ratios (100:0, 80:20, 60:40, 40:60, 20:80, and 0:100, on a volume basis) were conducted. Volatile solid (VS) concentration strongly affected the hydrogen production rate and yield compared with the waste mixing ratio. The specific hydrogen production rate was predicted to be optimal when the VS concentration ranged from 10 to 12 g/l at all the mixing ratios of molasses wastewater and sewage sludge. A hydrogen yield of over 50 ml $H_2/gVS_{removed}$ was obtained from mixed waste of 10% sewage sludge and 10 g/l VS (about 10-fold dilution ratio). The optimal chemical oxygen demand/total nitrogen ratio for co-digesting molasses wastewater and sewage sludge was between 250 and 300 with a hydrogen yield above 20 ml $H_2/gVS_{removed}$.

• Korean journal of applied entomology
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• 제44권3호
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• pp.189-197
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• 2005

Cellulose-digesting bacteria were isolated from hindgut of Allomyrina dichotoma (Coleoptera: Dynastidae). The bacterial isolates were identified as Yersinia sp. and Bacillus sp. The addition of the identified bacteria to diet increased growth rate of the cetoniid beetle, Protaetia brevitarsis senlensis (Coleoptera: Cetoniidae), probably by digesting cellulose nutrient contained in the oak tree sawdust diet. An additive of wheat flour at more than 10% to the sawdust diet significantly enhanced growth of P. brevitarsis senlensis. Trimmed branches of apple trees have been disposed in the apple farms and could be used for a diet component of the cetoniid beetle when the cellulose-digesting bacteria were mixed with the derived-sawdust. Resulting manure from mass rearing of P. brevitarsis senlensis contained high organic matters and trace amounts of toxic metals. When the manure were splayed on soil, it was effective as a natural compost and significantly stimulated lettuce growth. This research suggests a model technology to use cellulose-digesting bacteria to use for culturing grub, which results in natural recycles of trimmed branches in apple farms as grub diet, and to use grub manure as a natural compost.