• Journal of Microbiology and Biotechnology
• /
• v.25 no.1
• /
• pp.33-43
• /
• 2015

GAM-1 and GAM-2, two themostable glucoamylases from Aspergillus niger B-30, possess different molecular masses, glycosylation, and thermal stability. In the present study, the effects of additives on the thermal inactivation of GAM-1 and GAM-2 were investigated. The half-lives of GAM-1 and GAM-2 at 70℃ were 45 and 216 min, respectively. Data obtained from fluorescence spectroscopy, circular dichroism spectroscopy, UV absorption spectroscopy, and dynamic light scattering demonstrated that during the thermal inactivation progress, combined with the loss of the helical structure and a majority of the tertiary structure, tryptophan residues were partially exposed and further led to glucoamylases aggregating. The thermal stability of GAM-1 and GAM-2 was largely improved in the presence of sorbitol and trehalose. Results from spectroscopy and Native-PAGE confirmed that sorbitol and trehalose maintained the native state of glucoamylases and prevented their thermal aggregation. The loss of hydrophobic bonding and helical structure was responsible for the decrease of glucoamylase activity. Additionally, sorbitol and trehalose significantly increased the substrate affinity and catalytic efficiency of the two glucoamylases. Our results display an insight into the thermal inactivation of glucoamylases and provide an important base for industrial applications of the thermally stable glucoamylases.

• Microbiology and Biotechnology Letters
• /
• v.24 no.5
• /
• pp.604-612
• /
• 1996

To improve the fermentation characteristics of the haploid starch-fermenting recombinant yeast strain K114/YIpMS$\Delta$R(LEU2/URA3) secreting both $\alpha$-amylase and glucoamylase was rare-mated with polyploid industrial yeast Saccharomyces sp. K35. The K35 strain had good fermentation-characteristics such as ethanol-tolerance, high temperature and sugar-tolerance, and high fermentation rate. Among the resulting 66 hybrids, the best strain RH51 was selected. The RH51 exhibited amylolytic activity of K114/YIpMS$\Delta$R(LEU2/URA3) as well as ethanol and sugar tolerance of K35. The optimum temperature of hybrid RH51 for starch fermentation was 34$\circ$C which was same as that of K35 but different from that (30$\circ$C) of K114/YIpMS$\Delta$R(LEU2/URA3). The optimum pH was 5.0. The optimum size of inoculum was 2% as the pellet (w/v) of yeast cells. The hybrid strain RH51 produced 7.0% ethanol (w/v) from 20% (w/v) soluble starch while K35 formed almost no ethanol, 0.3% (w/v). RH51 strain produced 7.5% (w/v) ethanol after 8 days in a 2.5 l fermenter containing 800 ml of 20% (w/v) soluble starch. The residual starch content in the fermentation medium was 1.68% (w/v), and therefore almost all the starch was fermented completely.

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

• Preventive Nutrition and Food Science
• /
• v.20 no.3
• /
• pp.210-214
• /
• 2015

Grape pomace is an abundant source of underutilized winery by-products. Polyphenols were extracted from grape pomace using cellulase and gluco-amylase enzymes. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin-Ciocalteu's assays were used to measure antioxidant activity and total polyphenolic contents. Both cellulase, and gluco-amylase digested grape pomace showed efficient radical scavenging activity. In addition, the total polyphenolic contents of cellulase digested grape pomace showed lower concentrations were effective compared to higher concentrations, whereas glucoamylase enzyme did not show remarkable variations. The DPPH radical scavenging activity and total polyphenolic contents were significantly higher in the cellulase digested grape pomace compared to the gluco-amylase digested and the not digested grape pomace. It is notable that enzymatic digestions were efficient for extracting polyphenols from grape pomace. The underutilized grape pomace polyphenols can be further used for food safety as a natural antioxidant.

• Journal of the Korean Society of Tobacco Science
• /
• v.9 no.2
• /
• pp.69-75
• /
• 1987

Nicotine, the main alkaloid of tobacco, showed different effect according to the enzyme. Among investigated enzymes, protease was inactivated remarkably by nicotine and the mode of inhibition was examined. $\alpha$-amylase and $\beta$-amylase were not affected, and cellulase and glucoamylase were inactivated partially when the concentration of it was over 1.0% , but protease was inhibited powerfully by nicotine The inhibition of protease by nicotine was performed almost in the initial stage of reaction, and was not so much affected by temperature, and was reversible. The inhibition type of protease by nicotine appeared as a Mixed-type inhibition.

• Journal of Life Science
• /
• v.11 no.4
• /
• pp.304-310
• /
• 2001

In order to produce a high quality and functional black bean Chungkugjang, a bacterium which has potent enzyme activities(protease:124.8 U/$m\ell$, $\alpha$-amylase: 78.2U/$m\ell$, glucoamylase; 13.9U/$m\ell$, ), was isolated by using the halo zone method and identified by morphological, cultural and biochemical properties, and then finally confirmed by GP-microplate identification system as Bacillus megatrium SMY-212. The isolated SMY-212 system was also high in the formation of mucoid material and fibrinolytic activity.

• Korean Journal of Food Science and Technology
• /
• v.33 no.3
• /
• pp.288-293
• /
• 2001

${\alpha}-amylase$ activities of bean water was not significantly influenced by bean water concentrations but they were remarkably influenced by different temperatures and substrates. ${\alpha}-amylase$ activities of bean water on cooked starch were significantly higher than those on raw starch. ${\beta}-amylase$ and glucoamylase activities in 14% bean water were significantly higher than those in 7% bean water. Yukwa paste is glutinous rice flour paste. Bean water was added to Yukwa paste by 0, 7, 14% and incubated 0, 3, 6, 9, 12 hours at $60^{\circ}C$. The pH of Yukwa paste increased with bean water concentration and decreased with the incubation time. The viscosity decreased with bean water concentration and incubation time. The ruducing sugar content of Yukwa paste increased with bean water concentration and incubation time. The changes of reducing sugar content in cooked Yukwa paste were much higher than those in the raw one. ${\alpha},\;{\beta}-amylase$glucoamylase activities of Yukwa paste also increased with bean water concentration, and their activities were much higher on the cooked glutinous rice flour than those on the raw one. The SEM observation on the freeze-dried flour of Yukwa paste showed breakdown of amylopectin structure by addition of bean water in the paste.

• The Korean Journal of Mycology
• /
• v.51 no.1
• /
• pp.39-49
• /
• 2023

This study aimed to screen and investigate the microbiological characteristics, enzyme activity, and physiological functionality of unrecorded wild yeasts from the Wolsung valley of South Deogyu mountain and Jinjamcheon in Daejeon city, Korea. Candida sorboxylosa WSC25-4 (NNIBRFG47336), Arxula adeninivorans WSC6-3 (NNIBRFG47335), Farysizyma taiwaniana JRC6-2 (NNIBRFG47339), Saturnispora dispora JTS19-1 (NNIBRFG47334), Vanderwaltozyma polyspora JJCH5-3 (NNIBRFG47337), and Vanrija fragicola SW-9 (NNIBRFG47338) have not been previously recorded. None of these formed spores or pseudo-mycelium, and four strains, including C. sorboxylosa WSC25-4, were global in shaped. Except for V. polyspora JJCH5-3, all strains grew well in the yeast extract-peptone-dextrose (YPD), potato-dextrose and yeast malt extract media. A. adeninivorans WSC6-3 and S. dispora JTS19-1 grew well in 40% glucose-containing YPD medium and V. fragicola SW-9 in 5% NaCl-containing YPD medium. The supernatant of A. adeninivorans WSC6-3, F. taiwaniana JRC6-2, and V. fragicola SW-9 exhibited 58.0, 59.0, and 59.0% of glucoamylase activity, respectively. C. sorboxylosa WSC25-4 also produced extracellular alkaline protease. Antioxidant activity of the supernatant from F. taiwanian JRC6-2 was high (79.8%), while the other five unrecorded yeasts demonstrated elevated antiaging superoxide dismutase (SOD)-like activity (99%). Anti-gout xanthin oxidase inhibitory activity of S. dispora JTS19-1 and V. fragicola SW-9 was also elevated at 79 and 80%, respectively.

• Korean Journal of Food Science and Technology
• /
• v.34 no.2
• /
• pp.269-273
• /
• 2002

The amlyolytic enzymes $({\alpha}-amlyase,\;{\beta}-amlyase,\;glucoamlyase)$ and protease activities were studied during Kimchi fermentation. The optimum pH of Kimchi was 4.1 within 2 days at $20^{\circ}C$. The optimum acidity calculated as lactic acid was 0.44% within 2 days at $20^{\circ}C$. On the first day of fermentation, ${\alpha}-amlyase$ activity was reduced from 0.49 unit/mg protein to 0.20 unit/mg protein but increased in the later stage of fermentation. In case of ${\beta}-amlyase,\;glucoamlyase$ and protease showed the highest activity of 505.73, 13.43 and 1.72 unit/mg protein at the 2nd day of fermentation at $20^{\circ}C$. In the sensory evaluation of Kimchi were estimated taste, color, texture and overall acceptability. Overall acceptability of kimchi showed the highest score value on the 2nd day of fermentation, respectively.

• Journal of Biomedical and Translational Research
• /
• v.19 no.4
• /
• pp.103-109
• /
• 2018

Aspergillus luchuensis 34-1 was inoculated into wheat pellets with different conditions of raw materials to produce nuruk. The degree of substrate reactivity improvement of steam treated raw materials compared with that of non-heat treated was analyzed. The water content of the pellet was adjusted to 25% and 35%, and steam treatment for 10 minutes improved the substrate reactivity at 2.1-fold and 3.1-fold, and sterilization was also possible. The characteristics of improvement pellet nuruk were investigated according to the degree of crushing and water content of raw materials according to the temperatures and humidities ($23^{\circ}C$, $30^{\circ}C$ and RH 60%, RH 80%). The pH of the pellet nuruk was higher depending on the temperature, humidity and moisture content of the koji were lower, and the pH of the flour-pellet nuruk was lower than that of 2 mm milling wheat-pellet nuruk according to milling degree. It can be seen that the milling degree affects the growth of mold. The acidity and amino acid were generally higher as fermentation time increased. Also, the higher the incubation temperature, humidity and moisture content, the higher the value. Glucoamylase activity was significantly the highest in moisture content 35% D2b nuruk, cultured at $30^{\circ}C$ and 80% RH for 38 hours. This is higher than the previous reports on glucoamylase of rice-koji or commercial nuruk using fungi isolated from traditional nuruk. From these study, it is expected that making of improvement pellet nuruk would save the fermentation time considerably compared with traditional nuruks.