• Journal of Microbiology and Biotechnology
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• v.20 no.2
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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 Microbiology and Biotechnology
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• v.9 no.5
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• pp.668-671
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• 1999

The gene encoding Schwanniomyces occidentalis $\alpha-amylase$(AMY) was introduced into Saccharomyces cerevisiae var. diastaticus which secreted only glucoamylase, by using a linearized yeast integrating vector to develop stable strains with a capability of secreting $\alpha-amylase$and glucoamylase simultaneously. A dominant selectable marker, the geneticin(G418) resistance gene (Gt^r$), was cloned into a vector to screen wild-type diploid transformants harboring the AMY gene. The amylolytic activities of transformants were about 3-7 times higher than those of the recipient strains. When grown in nonselective media, the transformants with the linearized integrating vector containing the AMY gene exhibited almost all of the mitotic stability after 100 generations.

• Journal of Ginseng Research
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• v.9 no.2
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• pp.270-284
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• 1985

In order to investigate the change of physicochemical properties of ginseng root starch during storage and heat treatment, the roots were stored for 15 days at 5 $^{\circ}C$, 15 $^{\circ}C$, 3$0^{\circ}C$ and 45$^{\circ}C$, and heated for 15 hours at 6$0^{\circ}C$, 7$0^{\circ}C$, 8$0^{\circ}C$, 9$0^{\circ}C$, respectively. The starch content was decreased from about 40% to 23-26% and sucrose content was increased from 4% to 12-16% during storage for 15 days at 5-45$^{\circ}C$. Maltose, which was not detected in fresh samples, was increased up to 8.5% during storage or heat treatment. Granular size of the starch was decreased and some of the granules were broken during storage. Amylose content in the starch was decreased from 33% to 20%, and blue value and alkali number of the starch were increased slightly, and solubility and swelling power of the starch were decreased during storage. 3 The higher storage temperature and the longer storage time, the starch was more susceptible to gelatinize, and the viscosity of the starch was lowered with the susceptibility of gelatinization. The susceptibility of degradation of the starch by the amylase was increased and amylolytic activities in ginseng root were, also, increased during storage.

• The Korean Journal of Food And Nutrition
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• v.12 no.3
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• pp.219-225
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• 1999

In order to improve the quality of nuruk a Korean traditional fermenting agent for brewing nuruk was modified by a new method inoculating Rhizopus japonicus T2 Aspergillus oryzae L2 and hansenula sp. BC26 which had been isolated from traditional Nuruk. And the characteristics of modified Nuruk were investimgated as compared with current fermenting agents such as commercial Nuruk and rice koji of As-pergillus kawachii. The odor of modified Nuruk was savory but those of commercial Nuruk and rice koji were fetid andinodorous repectively. The extracted waters of modified Nuruk commercial Nuruk and rice koji were yellow pale yellow and colorless and showed 6.15, 6.01 and 3.30 of pH respectively. Mod-ified Nuruk had 7.6${\times}$106CFU/g of yeast but commercial uruk and rice koji had no yeast. Commercial Nuruk had 1.0${\times}$102CFU/g of lactic acid bacteria but modified Nuruk and rice koji had no lactic acid bacteria. The amylolytic and proteolytic activities of modified Nuruk were much higher than those of commercial Nuruk or rice koji. Seed mash of modified Nuruk had ester aroma but that of commercial Nuruk rather offensive odor and that of rice koji neither ester aroma or offensive odor. It seemed that if the modified Nuruk is used in seed mashing the supplement of acidulant is need to lower pH. The quality of modified Nuruk was thought to be much better than that of commercial Nuruk or rice koji.

• Microbiology and Biotechnology Letters
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• v.3 no.2
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• pp.73-82
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• 1975

In the course of studies on the production of thermostable amylases by thermophilic actinomycetes isolated from soils the investigation was carried out on the production of $\alpha$-amylase by G-1011 strain which had presented the most remarkable $\alpha$-amylase formation ability among 128 amylolytic isolates. The results were as follows ： 1. Characteristics of G-1011 strain were compared with those descriptions of thermophilic actinomycetes given in Bergey's Manual. The strain was identical to these species of actinomycetes. The details of physiological properties of the strain luould he published in near future. 2. The optimum temperature for incubation of the cell growth of G-1011 strain and $\alpha$-amylase production by the strain was revealed to 5$0^{\circ}C$. 3. The effective medium for $\alpha$-amylase formation by the strain was consisted of 3.0%, soluble starch, 1.0%, peptone, 0.5%, yeast extract, 0.5%, NaCl, 0.1%, MgSO$_4$ㆍ7$H_2O$ 0.02%, $K_2$MPO$_4$and 0.002% FeSO$_4$ㆍ7$H_2O$. The pH of the medium was ajusted to 7.0 with phosphate buffer solution. 4. The maximum production of $\alpha$-amylase (3420 D. U/ml) by G-1011 strain resulted when it was grown for 16 hours with the culture of reciprocal shaking.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.137-144
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• 1985

Studies were conducted on the conditions for preparation of yeast protoplasts utilizing Hansenula anomala var. anomala FRI YO-32 as well as Saccharomyces cerevisiae KFCC 32356 and a lytic enzyme from the snail Helix pomatia. The cell wails of the strain FRI YO-32 and S cerevisiae were found to be resistant to activity of the snail lytic enzyme if they were not treated with thiol compounds. Dithiothreitol was found to be more effective than 2-mercaptoethanol, but the latter was considered to be practical. As factors influencing the formation of yeast protoplast, it was considered to be concentration and incubation time of 2-mercaptoethanol or the lytic enzyme, growth stages in yeast cultivation, initial number of yeast cells, and concentration of osmotic stabilizer (KCI). Optimum conditions for the preparation of yeast protoplasts were determined.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.221-224
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• 1989

Amylolytic filamentous fungus, Aspergillus oryzae and nonamylolytic sugar fermentable yeast, Saccharomyces cerevisiae were fused by protoplast fusion in order to develope microorganisms having their intergrated function. Aminoacid auxotrophic properties were used as a genetic marker of protoplast fusion, and 35% PEG 4000 was used as a fusogenic agent. Complementation frequengy of fusion was $4.6\times 10^{-6}$ Obtained fusants showed the morphology of yeast strains, the amylase activity and the ethanol productivity. Among the properties of the fusants, morphology and prototrophic property were sustained stably but their ethanol productivity from starch was reduced. Although fusant strains had 0.5-fold ethanol productivity compared to that of S. cerevisiae in glucose medium, they produced ethanol from strach by direct fermentation.

• Mycobiology
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• v.50 no.2
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• pp.132-141
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• 2022

Amylomyces rouxii is commonly found as amylolytic fungi in tapai fermentation. However, its diversity is rarely reported despite being often used for food production in Southeast Asia. This research aims to analyze the genetic diversity and the distribution pattern of A. rouxii from Ragi tapai in Java Island, Indonesia. We isolated the fungus from samples obtained from Ragi tapai producing centers in Bandung, Sumedang, Muntilan, Blora, Yogyakarta, and Bondowoso. The obtained isolates were molecularly identified based on the ribosomal regions ITS1/ITS2 and D1/D2, then analyzed for phylogenetic tree reconstruction, genetic distance, genetic variation, and haplotype networking. Six isolates showed specific morphological traits of A. rouxii. However, phylogenetic tree reconstruction on the ribosomal genes showed that the isolates were grouped into two different clades related to two species. Clade A included BDG, SMD, and MTL isolates related to A. rouxii, whereas clade B included YOG, BLR, and BDS isolates related to Mucor indicus. The genetic distances between clades for ITS1/ITS2 and D1/D2 were 0.6145 and 0.1556, respectively. In conclusion, we confirmed the genetic diversity of molds from Ragi tapai in Java Island and showed that the isolates are not only related to A. rouxii as reported before.

• Journal of Ecology and Environment
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• v.31 no.2
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• pp.131-137
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• 2008

This study investigated the population densities and diversity of heterotrophic bacteria, and the rhizosphere-to-soil ratios (R/S) in the rhizosphere soil of halophyte Phragmites communis at the western coastal mudflats of Korea. The population densities of aerobic heterotrophic bacteria on the rhizosphere soil of P. communis were in the range of $3.3\;{\pm}\;0.9\;{\times}\;10^7\;{\sim}\;1.2\;{\pm}\;0.5\;{\times}\;10^8\;cfu\;g^{-1}$ dry weight (d. wt.). Population densities of amylolytic bacteria ranged from $1.1\;{\pm}\;0.2\;{\times}\;10^6$ to $3.0\;{\pm}\;1.2\;{\times}\;10^6\;cfu\;g^{-1}\;d.\;wt.$, while those of cellulolytic bacteria and proteolytic bacteria ranged from $5.6\;{\pm}\;2.3\;{\times}\;10^6$ to $1.5\;{\pm}\;0.3\;{\times}\;10^7\;cfu\;g^{-1}\;d.\;wt.$ and from $1.4\;{\pm}\;0.3\;{\times}\;10^6$ to $3.5\;{\pm}\;2.3\;{\times}\;10^7 \;cfu\;g^{-1}\;d.\;wt.$, respectively. The R/S ratios ranged from 2.26 to 6.89. Genetic (16S DNA) analysis of fifty-one isolates from the roots of P. communis suggested that the dominant species were closely related to the ${\gamma}$-proteobacteria group (18 clones) and the ${\alpha}$-proteobacteria group (14 clones). We found that halophyte species and mudflat environment both affected the rhizosphere bacterial communities.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.457-464
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• 2008

An extracellular enzyme (RMEBE) possessing ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferring activity was purified to homogeneity from Rhodothermus marin us by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex-200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at $80^{\circ}C$. Its half-life was determined to be 73.7 and 16.7 min at 80 and $85^{\circ}C$, respectively. The enzyme was also halophilic and highly halotolerant up to about 2M NaCl, with a maximal activity at 0.5M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial ${\alpha}-(1{\rightarrow}4)$-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferase activity for small maltooligosaccharides, producing disproportionated ${\alpha}-(1{\rightarrow}6)$-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity.