• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1521-1526
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• 2007

A maltogenic amylase gene from Lactobacillus gasseri ATCC33323 (LGMA) was expressed in Lactococcus lactis MG1363 using the P170 expression system. The successful production of recombinant LGMA (rLGMA) was confirmed by the catalytic activity of the enzyme in liquid and solid media. The N-terminal amino acid sequencing analysis of the rLGMA showed that it was Met-Gln-Leu-Ala-Ala-Leu-, which was the same as that of genuine protein, meaning the signal peptide was efficiently cleaved during secretion to the extracellular milieu. The optimal reaction temperature and pH of rLGMA ($55^{\circ}C$ and pH 5, respectively) and enzymatic hydrolysis patterns on various substrates (${\beta}$-cyclodextrin, starch, and pullulan) supported that rLGMA was not only efficiently secreted from the Lactococcus lactis MG1363 but was also functionally active. Finally, the branched maltooligosaccharides were effectively produced from liquefied com starch, by using rLGMA secreted from Lactococcus lactis, with a yield of 53.1%.

• Journal of Microbiology
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• v.42 no.4
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• pp.319-327
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• 2004

An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.815-818
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• 2003

Maltosyl(G2)-erythritol, produced by the transglycosylation reaction of erythritol with maltotriose by Bacillus stearothermophilus maltogenic amylase, was not utilized either as a substrate for lactic acid production or for water-insoluble glucan synthesis. An inhibition assay of dextransucrase and mutansucrase showed that the dental caries suppression effect of G2-erythritol was greater than that of erythritol.

• Proceedings of the Korean Society of Life Science Conference
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• 2003.05a
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• pp.23-37
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• 2003

A novel inhibitor against maltose-producing a-amylase was prepared via stepwise degradation of a high molecular weight acarbose (HMWA) using Thermus maltogenic amylase (ThMA). The structure of the purified inhibitor was determined to be ${\alpha}$-D-glucopyranosyl-${\alpha}$-acarviosinyl-D-glucopyranose (GlcAcvGlc). Progress curves of p-nitrophenyl-${\alpha}$-D-maltoside (PNPG2) hydrolysis by various amylolytic enzymes, including maltogenase (MGase), ThMA, and cyclodextrinase(CDase) I-5, in the presence of acarbose or GlcAcvGlc indicated a slow-binding mode of inhibition. The inhibition potency of GlcAcvGlc for MGase, ThMA, and CDase I-5 was 3 orders of magnitude higher than that of acarbose.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1093-1100
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• 2015

Acetate and lactate in growth media are detrimental to the production of Thermus maltogenic amylase (ThMA), a heterologous protein, as well as to the growth of recombinant Escherichia coli. Only 50 mM of acetate or 10 mM of lactate reduced 90% of specific ThMA activity. In this study, mutant E. coli strains blocked in the ackA-pta or ackA-pta and ldh pathways were created, characterized, and assessed for their culture performace in 300 L-scale fermentation. The ackApta and ldh double-mutant strain formed significantly less lactate and acetate, and produced a concomitant increase in the excretion of pyruvate (17.8 mM) under anaerobic conditions. The ackA-pta mutant strain accumulated significant acetate but had an approximately 2-fold increase in the formation of lactate. The ackA-pta and ldh double-mutant strain had superior overall performance in large-scale culture under suboptimal conditions, giving 67% higher cell density and 66% higher ThMA activity compared with those of the control strain. The doublemutant strain also achieved a 179% improvement in volumetric ThMA production.

• BMB Reports
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• v.35 no.6
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• pp.568-575
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• 2002

An $\alpha$-amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipiation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and $60^{\circ}C$. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had $\alpha$-anomeric forms, as determined by $^1H$-NMR. This maltose-forming $\alpha$-amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62%, repectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.

• Proceedings of the Korean Biophysical Society Conference
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• 1999.06a
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• pp.35-35
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• 1999

Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. The structure and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N -terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (${\alpha}$/ ${\beta}$)$\sub$8/ barrel of the adjacent subunit.(omitted)

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.761-766
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• 2008

The effects of various enzymes and emulsifiers on the loaf volume and crumb hardness of rice breads were studied. Four different enzymes [fungal ${\alpha}$-amylase (AMYL), maltogenic bacterial ${\alpha}$-amylase (NMYL), glucose oxidases (GO), and xylanase+hemicellulases (PTP)] and four emulsifiers [sorbitan monostearate (SMS), glycerol monostearate (GMS), sodium stearoyl lactylate (SSL), and glycerol ester+propylene glycol ester+sucrose ester+sorbitan ester (SP)] were supplemented to rice dough. The addition of AMYL, GO, and GO+AMYL increased loaf volume of rice breads. The highest loaf volume was observed in rice bread supplemented with AMYL. Rice breads supplemented with enzymes firmed at lower rates during storage, and AMYL, NMYL, and GO considerably decreased crumb hardness of rice breads, exhibiting a significant antistaling effect. The addition of emulsifiers produced rice breads with better specific loaf volume and crumb texture, and continuously retarded crumb hardness of rice breads during storage. Especially, rice bread supplemented with SSL demonstrated the highest loaf volume and the lowest crumb hardness during storage.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.851-855
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• 2006

Acarviosine-glucose (AcvGlc) is an ${\alpha}$-glucosidase inhibitor and has similar inhibitory activity to acarbose in vitro. We synthesized AcvGlc by treating acarbose with Bacillus stearothermophilus maltogenic amylase and fed C57BL/6J and db/db mice with diets containing purified AcvGlc and acarbose for 1 week. AcvGlc (50 and 100 mg/100 g diet) significantly reduced plasma glucose and triglyceride levels in db/db mice by 42 and 51 %, respectively (p<0.0001). The hypoglycemic and hypotriglyceridemic effects of AcvGlc were slightly, but significantly, greater than those seen with acarbose treatment (p<0.0001) in C57BL/6J mice. In an oral glucose tolerance test, glucose tolerance was significantly improved at all time points (p<0.01). The expression of two novel glucose transporters (GLUTs), GLUT10 and GLUT12, were examined by Western blot analysis. GLUT10 was markedly increased in the db/db livers. After AcvGlc treatment, the expression of hepatic GLUT10 was decreased whereas intestinal GLUT12 was significantly increased in both strains of mice. Our results show that AcvGlc improves plasma lipid and glucose metabolism slightly more than acarbose. Regulation of hepatic GLUT10 and intestinal GLUT12 may be important in controlling blood glucose levels.

• Food Engineering Progress
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• v.14 no.3
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• pp.235-242
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• 2010

This study was to investigate the combined effect of enzyme and trehalose addition on the retrogradation rate and quality changes in a Korean traditional rice cake, Backsulgi. As for the enzyme, a commercial maltogenic amylase, Novamyl, was used. From texture profile analysis and sensory tests, the optimum enzyme content for the preparation of Backsulgi was determined to be 0.1%. Backsulgi was prepared with the fixed content of Novamyl (0.1%) and different ratios (5, 10, 15%) of trehalose, and physicochemical and sensory properties were examined for 3 days of storage at ${25^{\circ}C}$. As trehalose content increased, the water activity of Backsulgi decreased. In Hunter's color value, L- and a-values of trehalose-added samples were significantly lower than those of control. In texture profile analysis, hardness of Backsulgi decreased with trehalose content. During storage, a increase in hardness and a decrease in cohesiveness were typically observed for all samples. However, those changes were significantly reduced with trehalose content. The retardation of retrogradation with the enzyme and trehalose addition was confirmed using calculated Avrami rate and time constants. These results revealed that combined treatments of 0.1% Novamyl and 10% trehalose could produce Backsulgi of better sensory quality with retarded retrogradation compared to separated treatment.