• Journal of Microbiology and Biotechnology
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• 제17권9호
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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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• 제42권4호
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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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• 제13권5호
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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.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2003년도 제39회 학술심포지움
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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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• 제25권7호
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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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• 제35권6호
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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.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1999년도 학술발표회 진행표 및 논문초록
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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)

• 한국식품영양과학회지
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• 제37권6호
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• pp.761-766
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• 2008

효소제와 유화제의 첨가가 쌀빵의 품질특성중 빵의 부피와 저장중 경도 변화에 미치는 영향을 조사하였다. 효소제로는 곰팡이 ${\alpha}$-amylase(AMYL), 세균 ${\alpha}$-amylase(NMYL), glucose oxidase(GO), 그리고 xylanase+hemicellulase (PTP)를 사용하였으며, 유화제로 sorbitan monostearate (SMS), glycerol monostearate(GMS), sodium stearoyl lactylate(SSL), glycerol ester+propylene glycol ester+sucrose ester+sorbitan ester(SP)를 첨가하여 그 효과를 비교하였다. 쌀가루에 AMYL, GO, 그리고 AMYL+GO 효소를 처리한 쌀빵의 비용적이 대조구 쌀빵에 비해 높게 나타났는데, 특히 AMYL을 첨가한 처리구에서 가장 높은 수치를 보여주었다. 쌀빵의 저장 중 경도 변화를 측정하였을 때 AMYL, NMYL, 그리고 GO를 첨가한 처리구에서 3일 경과 후에도 낮은 증가율을 나타내 쌀빵의 노화지연에 긍정적인 효과를 제시해 주었다. 쌀가루에 유화제를 첨가한 쌀빵의 부피는 모든 유화제 처리구가 대조구 쌀빵보다 높게 나타났으며 저장중 경도가 지속적으로 낮았다. 특히 SSL은 쌀빵의 부피 개선과 함께 노화지연의 효과가 가장 높아 쌀빵의 품질 개량을 위해 적용할 수 있는 매우 긍정적인 유화제로 평가되었다.

• Food Science and Biotechnology
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• 제15권6호
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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.

• 산업식품공학
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• 제14권3호
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• pp.235-242
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• 2010

본 연구는 백설기의 노화를 억제 및 저장 중 품질 향상을 위한 Novamyl의 최적 첨가량을 0.1%로 선정하고 트레할로스를 5%, 10%, 15% 첨가하여 백설기를 제조한 후 상 온(${25^{\circ}C}$)에서 0, 1, 2, 3일간 저장하면서 기계적 및 관능적 특성과 노화 속도 변화를 살펴보았으며 그 결과는 다음과 같다. 수분활성도는 트레할로스의 첨가량이 증가할수록 유의적으로 감소하였으며 저장 일수에 따른 차이는 없었다. 색도는 트레할로스를 첨가한 백설기의 명도(L값)가 유의적으로 낮게 나타났으며 적색도를 나타내는 a값은 첨가군이 대조구에 비해 낮게 나타났고 황색도(b값)는 첨가군이 다소 낮은 값을 보였으며 저장 일수가 증가함에 따라 황색도가 감소하는 경향을 보였다. 기계적 조직감에 대해서는 트레할로스의 첨가량이 증가할수록 경도가 감소하였고 저장 중 씹힘성의 감소폭이 감소하였다. Avrami 방정식으로 노화속도를 측정하였으며, Avrami exponent(n) 값과 시간상수(1/k) 값을 계산한 결과 트레할로스 첨가군이 대조구에 비해 노화속도가 감소하였음을 알 수 있었다. 관능검사 결과에서 15% 첨가구가 대부분의 항목에서 우수한 결과를 보였고 특히 전반적인 기호도 및 씹힘성 항목에서 높은 선호도 값을 보였으나 10% 첨가구와의 유의적인 차이는 보이지 않았다. 이와 같은 실험결과를 통해 백설기 제조 시 Novamyl 0.1%를 처리하고 트레할로스를 첨가할 경우 노화 억제와 품질 향상의 효과가 있었으며, 특히 기계적 측정 결과와 관능검사 결과를 종합했을 때, 트레할로스를 10% 첨가가 15%의 첨가에 비해 유의적인 차이가 적어 가장 적절한 것으로 나타났다. 효소의 첨가는 백설기의 노화를 억제하는 효과가 있지만 물성적인 측면에서 조직이 과도하게 물러지는 등의 문제를 보이는데 이에 트레할로스를 적당량 첨가함으로써 노화 억제를 향상시키고 물성적인 측면과 기호도를 향상시켜 저장성이 우수하고 소비자의 입맛에 적합한 백설기의 제조가 가능하다고 사료된다.