• Journal of Microbiology and Biotechnology
• /
• v.22 no.5
• /
• pp.592-599
• /
• 2012

Bacillus licheniformis ${\alpha}$-amylase (BLA), a thermophilic counterpart of Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), is an appropriate model for the design of stabilizing mutations in BAA. BLA has 10 more histidines than BAA. Considering this prominent difference, in the present study, three out of these positions (I34, Q67, and P407; located in the thermostability determinant 1 region and Ca-III binding site of BAA) were replaced with histidine in BAA, using the site-directed mutagenesis technique. The results showed that the thermostability of P407H and Q67H mutants had increased, but no significant changes were observed in their kinetic parameters compared to that of the wild type. I34H replacement resulted in complete loss of enzyme activity. Moreover, fluorescence and circular dichroism data indicated a more rigid structure for the P407H variant compared with that of the wild-type BAA. However, the flexibility of Q67H and I34H mutants increased in comparison with that of wild-type enzyme.

• BMB Reports
• /
• v.40 no.1
• /
• pp.29-35
• /
• 2007

During the past decade, many salivary parameters have been used to characterize disease states. Ghrelin (GAH) is recently-discovered peptide hormone secreted mainly from the stomach but also produced in a number of other tissues including salivary glands. The aim of this work was to examine the relationship between active (aGAH) and inactive (dGAH) ghrelin in the saliva and other salivary parameters in type II diabetic patients and healthy controls. Salivary parameters were assessed in a single measurement of unstimulated whole saliva from 20 obese and 20 non-obese type II diabetes patients, and in 22 healthy controls. Total protein and alpha-amylase were determined by colorimetric methods, and glucose by the glucose-oxidase method. Saliva aGAH and dGAH levels were measured using a commercial radioimmunoassay (RIA) kit. Salivary concentrations of aGAH and dGAH ghrelin were more markedly decreased in obese diabetic subjects than in the two other groups. Glucose and alpha-amylase levels were higher in diabetic subjects than in controls. Furthermore, there were correlations between GAH levels and BMI, and between GAH and blood pressure. However, there was no marked variability in saliva flow rates among the groups. These results indicate that measurement of salivary GAH and its relationship to other salivary parameters might help to provide insight into the role of ghrelin in diabetes.

• Preventive Nutrition and Food Science
• /
• v.3 no.1
• /
• pp.98-104
• /
• 1998

As an efffort ot construct LAB (latice acid bacteria), capable of utilizing starch as fermentation substrate without the aid of externally supplied enzymes, plasmid vectors containing the amyL($\alpha$-amylase/pullulansase gene) from Clostridium thermophydrosulfuricum, and glucoamylase cDNA from Asperigillus shirousamii were constructed and introduced itno E. coli and L. lactis. For expression in procaryotes , 1.9kb glucoamylase cDNA encoding the mature form of enzyme was PCR amplified and translationaly fused to a PCR amplified 260 bp fragment containing the promotor and secretion signals of amyl in the same reading frame. The production of $\alpha$-amylase, Apu, and glucoamlase in E. coli and L. lactis was confirmed by enzyme assay and zymography . Enzymeswere detected in both cellpellets and supernatants, indicating theworking of scretion signals in heterologous hosts. The efficiencies of secretion were varibale depending on the gene and host. The highest $\alpha$- amylase acitivity observed was 1.1 units and most activiity was detected from thecell pellets. The degree of gene expression in both hosts and the effect on the growth of hosts were examined.

• Preventive Nutrition and Food Science
• /
• v.6 no.4
• /
• pp.206-210
• /
• 2001

Starch is an abundant resource in plant biomass, and it should be hydrolyzed enzymatically into fermentable sugars for ethanol fermentation. A genetic recombinant yeast, Saccharomyces cerevisiae GA-7458, was constructed by integrating the structural gene of both $\alpha$-amylase from Bacillus stearothermophilus and the gene (STA1) encoding glucoamylase from S. diastaticus into the chromosome of S. cerevisiae SH7458. The recombinant yeast showed active enzymatic activities of $\alpha$-amylase and glucoamylase. The productivity of ethanol fermentation from the pH-controlled batch culture (pH 5.5) was 2.6 times greater than that of the pH-uncontrolled batch culture. Moreover, in a fed-batch culture, more ethanol was produced (13.2 g/L), and the production yield was 0.38 with 2% of corn starch. Importantly, the integrated plasmids were fully maintained during ethanol fermentation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.36 no.8
• /
• pp.989-994
• /
• 2007

This study was designed to investigate whether Sasa borealis leaves extracts (SLE) may inhibit yeast ${\alpha}-glucosidase$ and ${\alpha}-amylase$ activities and postprandial hyperglycemia in STZ-induced diabetic mice. Freeze-dried SLE was extracted with 70% methanol and followed by a sequential fractionation with dicholoromethan, ethylacetate, butanol, and water. Both ethylacetate and butanol fractions showed high inhibitory activities against the ${\alpha}-glucosidase$ and ${\alpha}-amylase$ enzymes. The $IC_{50}$ of ethylacetate and butanol fractions against ${\alpha}-glucosidase$ were 0.54 and 0.63 mg/mL, respectively, indicating a greater inhibition effect than acarbose (0.68 mg/mL) (p<0.05). Likewise, the two fractions exhibited a smaller $IC_{50}$ against ${\alpha}-amylase$, compared with acarbose (p<0.05). However, the yield of ethylacetate fraction of SLE was relatively small. Postprandial blood glucose testing of normal mice and STZ-induced diabetic mice by starch soln. loading (2 g/kg B.W.) showed that postprandial blood glucose level at 30, 60, and 120 min were markedly decreased by single oral administration of SLE butanol fraction (200 mg/kg B.W.) in both normal (p<0.0l) and diabetic mice (p<0.0l). Furthermore, the incremental area under the curve (AUC) was significantly lowered via SLE administration (5,745 versus 12,435 $mg{\cdot}mim/dL$) in the diabetic mice (p<0.0l). The incremental AUC in normal mice corroborated the hypoglycemic effect of SLE (p<0.0l) found in the diabetic mice. These results suggest that SLE may delay carbohydrate digestion and thus glucose absorption. In addition, SLE may have the potential to prevent and treat diabetes via its ability on lowering postprandial hyperglycemia.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.9
• /
• pp.1439-1445
• /
• 2013

The aim of this study was to investigate the quality characteristics of rice cookies specifically prepared with rice flour following enzyme treatment. In this study, three types of specific enzyme treatments, that is, ${\alpha}$-amylase, ${\beta}$-amylase, and cellulose+${\beta}$-glucanase were applied on the effect of rice flour with respect to desired quality characteristics of the rice cookies. Based on our study results, we have found that the density of dough in the rice cookies was not significantly different between that of the non-enzyme treated (control) and the enzyme treated group. However, we found that the spread factor of rice cookies, prepared with rice flour treated with ${\beta}$-amylase, was higher than that of the control. It was also found that the moisture content of rice cookies (with added enzyme-treated rice flour) was 3.20 to 3.90%; however, this range is much lower than that observed in the control. Further, we observed that Hunter color's L-values were significantly higher for the control than those of the enzyme-treated cookies. We also found that hardness of enzyme-treated cookies was comparatively better than that of the control. In addition, the sensory acceptability scores of the enzyme-treated cookies were found to be significantly higher than the control in decisive parameters such as aroma, appearance, flavor, taste, texture, and overall acceptability. Based on our findings, we suggest that ${\alpha}$-amylase treated rice flour is an effective ingredient for improving the overall quality of rice cookies.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.1 no.2
• /
• pp.123-129
• /
• 2000

Expression of the crylAcl gene of an acrystalliferous Bacillus thuringiensis strain under the control of the native or $\alpha$-amylase gene promoter was investigated. The crylAcl gene was cloned in a B. thuringiensis - E. coli shutle vector, pHT3101, undder the control of either the native promoter (pProAc) or the $\alpha$-amylase promoter from Bacillus subtilis (pAmyAc). These two recombinant plasmids were successfully expressed in B. thuringiensis subsp. kurstaki Cry B. The first transformant (ProAc/CB), harboring pProAc, expressed an about 130 kDa protein begining 24 hr after inoculations just as in the case of the wild type of B. thuringiensis subsp. kurstaki HD-73. The second pAmyAc-transformant (AmyAc/CB) began to express the gene just 6 hr after inoculation, but Western analysis showed that the activity of the $\alpha$-amylase promoter was relatively weaker than that of the native promoter. As expected, their toxicity against Plutella xylostella larvae was dependent on the amount of Cry1Acl protein expressed.

• Food Science and Biotechnology
• /
• v.17 no.2
• /
• pp.302-307
• /
• 2008

Novel amylase inhibitors were synthesized via transglycosylation by Thermotoga maritima glucosidase (TMG). TMG hydrolyzes acarbose, acarviosine-glucose, and maltooligosaccharide by releasing $^{14}C$-labeled glucose from the reducing end of each molecule. When TMG was incubated with acarviosine-glucose (the donor) and glucose (the acceptor), two major transfer products, compounds 1 and 2, were formed via transglycosylation. The structures of the transfer products were determined using thin-layer chromatography (TLC), high-performance ion chromatography (HPIC), and $^{13}C$ nuclear magnetic resonance (NMR) spectroscopy. The results indicate that acarviosine was transferred to glucose at either C-6, to give a $\alpha-(1{\rightarrow}6$) glycosidic linkage, or at C-3, to produce an $\alpha-(1{\rightarrow}3$) glycosidic linkage. The transfer products showed a mixed-type inhibition against porcine pancreatic $\alpha$-amylase; therefore, they may be useful not only as inhibitors but also as acarbose transition-state analogs to study the mechanism of amylase inhibition.

• Culinary science and hospitality research
• /
• v.23 no.1
• /
• pp.95-102
• /
• 2017

This study was activity of ${\alpha}-amylase$, glucoamylase of liquid Nuruk prepared using liquid Nuruk (NK) and Aspergillus kawachii (AK), Aspergillus niger (AN), Aspergillus oryzae (AO), Monascus kaoliang (MK). To investigate the relationship between the enzymatic activity and the total sugar content of liquid Nuruk depending on the types of Nuruk molds and the degree of rice-polishing. The activity of ${\alpha}-amylase$ depending on the types of Nuruk molds was shown to be 8.82, 8.72 units/mL in AN and AK treatments in brown rice liquid Nuruk at 24 hours after incubation, as the degree of rice-polishing increased, the activity of ${\alpha}-amylase$ was significantly lower (p<0.05). When brown rice was incubated in AN, it showed 8.83 units/mL at 48 hours after incubation, which was the highest activity, but there was no significantly difference (p<0.05), as the degree of rice- polishing was higher, the activity of ${\alpha}-amylase$ was lower. The activity of glucoamylase depending on the degree of rice-polishing showed 3,013 units/mL in AO treatment in brown rice liquid Nuruk at 24 hours after incubation, and the enzymatic activity was significantly higher (p<0.05). As the degree of rice-polishing increased, the activity of glucoamylase decreased, so liquid brown rice Nuruk showed the highest enzymatic activity, liquid white rice Nuruk was the lowest enzymatic activity. The highest enzymatic activity appeared in liquid Nuruk with brown rice at 48 hours after incubation. The activity of ${\alpha}-amylase$, glucoamylase showed higher enzymatic productivity as the degree of rice-polishing was lower, and there was an inverse correlation with the total sugar content.

• Korean Journal of Food Science and Technology
• /
• v.29 no.1
• /
• pp.101-106
• /
• 1997

This study was carried out to examine the possibility of using brown rice or paddy as raw materials for sikhe. Brown rice and paddy were soaked in water at $15^{\circ}C$ for 2 days and then germinated at $15,\;20,\;25,\;30^{\circ}C$ for upto 10 days. The higher the germination temperature, the higher the germination speed, the increase of ${\alpha}-amylase$ activity, and the increase of extract amount and its sugar content. The viscosity of extract rapidly decreased first and then slightly increased during germination. The activities of ${\alpha}-amylase$ of germinated brown rice and paddy were much smaller than those of germinated barley; however, the extract amount and its sugar content of germinated brown rice and paddy were similar to those of germinated barley. The germinated brown rice can be used for sikhe not as a saccharifying agent but as a substitute for white rice.