• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.231-237
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• 1988

Several methods to produce ethanol from tapioca starch were examined. Among four methods tested, alcohol fermentation with extruded tapioca starch was the most effective, which alcohol yield was 460.5 f/ton. After 69hours reaction with Rhizopus sp. glucoamylase, 108.7mg/$m\ell$ of reducing sugar were produced from extruded tapioca and 43.8mg/$m\ell$ from raw tapioca starch. In alcohol fermentation with extruded tapioca, the high concentration of alcohol at early stage prevented bacterial contamination and the fermentation rate was increased due to the high saccharifying power of glucoamylase on the extruded starch, but extrusion temperature had no influence on the fermentability, Scanning electron microscopy showed that the extrusion process changed the structure of tapioca starch granule to more susceptible form to glucoamylase attack than the raw starch. And glucoamylase of Rhizopus sp. had stronger digestion activity on both extruded tapioca and raw tapioca starch than that of Aspergillus usamii.

• Korean Journal of Agricultural Science
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• v.10 no.1
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• pp.166-185
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• 1983

Aspergillus niger IFO 8541 (NRRL 3112) was investigated through a series of UV rays and N-Methyl-N'-Nitro-N-Nitrosoguanidine (NTG) treatments to induce mutants that produce highly active raw starch saccharifying enzyme, and two mutants with strong enzymatic productivity were obtained. The mutants obtained were investigated for their fungal characters, condition of enzyme production, and other activities. Furthermore, the raw starch saccharifying enzyme was purified and the characteristics of purified enzyme were studied. The results obtained were summarized as follows; 1. The color of conidial head of UV-46 mutant obtained from UV rays treatment was changed to tan type and the gelatinated starch saccharifying enzyme productivity and the raw starch saccharifying enzyme productivity increased up to twice and 1.8 times compared to the productivities of original Aspergillus niger IFO 8541 cultured on the wheat bran, respectively. 2. The conidial head color of NG-41 mutant obtained from NTG treatment became lighter than that of parent strain. The gelatinated starch saccharifying enzyme productivity and raw starch saccharifying enzyme productivity increased about 1.8 times, and twice over the Aspergillus niger IFO 8541 parent strain cultured on wheat bran, respectively. The productivity of ${\alpha}$-amylase increased about 3 times more than the parent strain. 3. Two peaks of glucoanlylase and a peak of ${\alpha}$-amylase were obtained when enzyme solution of mutants and parent strain were passed through DEAE-Sephadex A-50 column chromatography. Glucoamylase I showed only gelatinated starch saccharifying enzyme activity. However, glucoamylase II (raw starch saccharifying enzyme) showed both raw starch saccharifying enzyme activity and gelatinated starch saccharifying enzyme activity. 4. Mutant, UV-46 was strengthened in glucoamylase II productivity and mutant NG-41 was strengthened in ${\alpha}$-amylase productivity. 5. Glucoamylase II of mutants and parent strain were appeared to have the same enzymatic properties. 6. Glucoamylase II of mutants and parent strain were recognized as simple enzyme through electrophoresis. 7. The glucoamylase II crystallized showed rhombic board type. 8. The molecular weight, isoelectric point, optimum pH, and optimum temperature of the glucoamylase II crystallized were estimated as 76,000, 3.4, 3.5 and $60^{\circ}C$, respectively.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1865-1875
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• 2018

Enhanced application of solid-state fermentation (SSF) in industrial production and the influence of SSF of Rhizopus K1 on glucoamylase productivity were analyzed using the flat band method. A growth model was implemented through SSF of Rhizopus K1 in this experiment, and spectrophotometric method was used to determine glucoamylase activity. Results showed that in bran and potato culture medium with 70% moisture in a loose state, ${\mu}$ of mycelium reached to $0.15h^{-1}$ after 45 h of culture in a thermostatic water bath incubator at $30^{\circ}C$. Under a low-magnification microscope, mycelial cells appeared uniform, bulky with numerous branches, and were not easily ruptured. The generated glucoamylase activity reached to 55 U/g (dry basis). This study has good utilization value for glucoamylase production by Rhizopus in SSF.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.185-195
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• 2015

Glucoamylase is an important industrial enzyme. Glucoamylase production by industrial Aspergillus niger strain featured with two major problems: (i) empirical substrate feeding methods deteriorating the fermentation performance; and (ii) the high raw materials cost limiting the economics of the glucoamylase product with delegated specification. In this study, we first proposed a novel three-stage varied-rate substrate feeding strategy for efficient glucoamylase production in a 5 L bioreactor using the standard feeding medium, by comparing the changing patterns of the important physiological parameters such as DO, OUR, RQ, etc., when using different substrate feeding strategies. With this strategy, the glucoamylase activity and productivity reached higher levels of 11,000 U/ml and 84.6 U/ml/h, respectively. The performance enhancement in this case was beneficial from the following results: DO and OUR could be controlled at the higher levels (30%, 43.83 mmol/l/h), while RQ was maintained at a stable/lower level of 0.60 simultaneously throughout the fed-batch phase. Based on this three-stage varied-rate substrate feeding strategy, we further evaluated the economics of using alternative carbon sources, attempting to reduce the raw materials cost. The results revealed that cornstarch hydrolysate could be considered as the best carbon source to replace the standard and expensive feeding medium. In this case, the production cost of the glucoamylase with delegated specification (5,000 U/ml) could be saved by more than 61% while the product quality be ensured simultaneously. The proposed strategy showed application potential in improving the economics of industrial glucoamylase production.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1690-1698
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• 2014

Partial purification of glucoamylase from solid-state fermentation culture was, firstly, investigated by reverse micellar extraction (RME). To avoid back extraction problems, the glucoamylase was kept in the original aqueous phase, while the other undesired proteins/enzymes were moved to the reverse micellar organic phase. The individual and interaction effects of main factors (i.e., pH and NaCl concentration in the aqueous phase, and concentration of sodium bis-2-ethyl-hexyl-sulfosuccinate (AOT) in the organic phase) were studied using response surface methodology. The optimum conditions for the maximum recovery of the enzyme were pH 2.75, 100 mM NaCl, and 200 mM AOT. Furthermore, the optimum organic to aqueous volume ratio ($V_{org}/V_{aq}$) and appropriate number of sequential extraction stages were 2 and 3, respectively. Finally, 60% of the undesired enzymes including proteases and xylanases were removed from the aqueous phase, while 140% of glucoamylase activity was recovered in the aqueous phase and the purification factor of glucoamylase was found to be 3.0-fold.

• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.497-503
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• 1991

The catalytic activities of immobilized gIucoamylase in a packed bed column and a continuous stirred tank reactor have been compared. Rapid production of glucose from liquefied starch have been studied through, the continuous liquefaction and saccharification using settling chamber. The immobilized glucoamylase with chitin gave the saccharification yield of 20% with the dextrin concentration of 100 g/l in a residence of 20 min. in a packed bed column. The half-life of immobilized glucoamylase with chitin was 19 days. The glucoamyalse immobilized in chitin and encapsulated with Ca-alginate gave the saccharification yield of 6% with the dextrin concentration of 50 g/l in a residence of 20 min. in a packed bed column. The Ca-alginate encapsulated and chitin immobilized glucomylase had a half-life of 25 days, which is 6 day larger than that of the immobilized glucoamylase with chitin only. In continuous liquefaction and saccharification, the glucose yield was 17% for the liquefied starch with naked barley concentration of 50gA in a residence of 20 min.

• BMB Reports
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• v.28 no.4
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• pp.342-347
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• 1995

The induction of glucoamylase biosynthesis from the yeast Candida tsukubaensis by different carbon sources was investigated by using either an enzyme activity assay or immunoblot analysis. The induction by C. tsukubaensis appears to be independent of the carbon sources, although the level of enzyme activity was lower in slowly utilizable carbon sources such as galactose. This glucoamylase is a constitutive enzyme and its biosynthesis is resistant to carbon catabolite repression. Glucose was more effective for the enzyme induction than starch, maltose or glycerol. In addition, this enzyme is regulated by both induction and repression.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.499-506
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• 1994

In order to convert starch to the fermentative sugar, the effect of pullulanase on the saccharification of starch and pullulanase was investigated. The optimum pH and temperature for the enzyme activity of the glucoamylase and the crude pullulanase from Klebsiella pneumoniae NFB-320 were shown to be identical as pH 6.0 and 60$\circC, respectively. The crude pullulanase was stable between pH 5.0~6.5, and up to 40$\circC, whereas the glucoamylase was stable between pH 4.0~6.5, and up to 40$\circC. When pullulanase and glucoamylase were engaged together in the sacchrification of starch, saccharification yield was increased by 3.2% than the yield obtained by glucoamylase, alone. And the two enzymes produced sugar from pulltulan 18 times much higher than the single use of pullulanase.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.320-326
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• 1988

Glucoamylase (EC 3.2.1.3) of Aspergillus sp. isolated from soil was partially purified by Sephacryl S-200 gel filtration and DEAE-Sephacel ion exchange chromatography, The glucoamylase activity was separated into two isozymes after DEAE-Sephacel ion exchange chromatrography. The optimum pH and temperature for both glucoamylase isozymes (GI, GII) were identical; pH 4.5 and temperature, $65^{\circ}C$. The molecular weights of GI and GII Isozymes were estimated to be 105,000, which were measured by gel filtration on Sephacryl S-200. Both isozymes were stable at pH ranges of 2 to 7, and up to 6$0^{\circ}C$. Glycerol was effective to stabilize the both isozymes. The activation energies of GI and GII isozymes were 10.63 and 10.33 kcal/mole, respectively. The enzyme activities of both isozymes were completely inactivated by addition of 0.1% Hg$^{++}$. In kinetic studies, the Km values of GI isozyme for soluble starch, dextrin, and glycogen were 0.62%, 0.32%, and 1.02%, respectively. For GII isozyme, they became 0.66%, 0.23%. and 0.14% for the substrates.

• Korean Journal of Microbiology
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• v.18 no.4
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• pp.161-171
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• 1980

A mutant strain having increased productivity of both enzymes, protease and amylase, was obtained from A. flavus KU 153, isolatd from South Korea for its high protease production by successive ultra-violet light irradiation, Two glucoamylases from the mutant strain selected were purified from wheat branculture by successive salting out, followed by dialysis and column chromatography, and their characteristics were compared with those of the wild strain. Glucoamylase production of the mutant selected was increased about 3.3 times compared with the wild strain, and 2.1 times compared with the parental strain, ${\alpha}-amylase$ activity of the mutant selected was about 2 times hugher than that of the wild strain or the parental strain. Protease and cellulase productivities of the muant selected were all alike compared with those of the highly proteolytic mutant, the parental strain. Therefore, it was considered that the back mutation on the protease production did not occurred in the formation process of the glucoamylase producing mutant. Total activities of glucoamylase I and II from the mutant selected were 2.86 and 3.65 times higher compared with those from the wild strain, respectively. Considering the optimal pH-thermal stability and Km-Vmax value of glucoamylase I and II from both strains, wild and mutant, it was deduced that the characteristics of glucoamylase I and II from the wild strain did not altered during the mutation process. Therefore, it was concluded that the selected mutant did not induce the formation of another glucoamylase isozyme, or the changes in the characteristics of the glucoamylase, but induce the productivity of the same glucoamylase I and II by the action of regulatory gene.