• Title/Summary/Keyword: $\alpha$-Amylase activity

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A New Raw-Starch-Digesting ${\alpha}$-Amylase: Production Under Solid-State Fermentation on Crude Millet and Biochemical Characterization

  • Maktouf, Sameh;Kamoun, Amel;Moulis, Claire;Remaud-Simeon, Magali;Ghribi, Dhouha;Chaabouni, Semia Ellouz
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.489-498
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    • 2013
  • A new Bacillus strain degrading starch, named Bacillus sp. UEB-S, was isolated from a southern Tunisian area. Amylase production using solid-state fermentation on millet, an inexpensive and available agro-resource, was investigated. Response surface methodology was applied to establish the relationship between enzyme production and four variables: inoculum size, moisture-to-millet ratio, temperature, and fermentation duration. The maximum enzyme activity recovered was 680 U/g of dry substrate when using $1.38{\times}10^9$ CFU/g as inoculation level, 5.6:1 (ml/g) as moisture ratio (86%), for 4 days of cultivation at $37^{\circ}C$, which was in perfect agreement with the predicted model value. Amylase was purified by Q-Sepharose anion-exchange and Sephacryl S-200 gel filtration chromatography with a 14-fold increase in specific activity. Its molecular mass was estimated at 130 kDa. The enzyme showed maximal activity at pH 5 and $70^{\circ}C$, and efficiently hydrolyzed starch to yield glucose and maltose as end products. The enzyme proved its efficiency for digesting raw cereal below gelatinization temperature and, hence, its potentiality to be used in industrial processes.

Studies on $\alpha$-amylase of Bocillus circulans F-2 (Part II) Enzymatic characteristics of the purified $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-amylase에 관한 연구 (제 I I 보) 정제$\alpha$-amylase의 효소적특성)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.123-132
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    • 1982
  • These experiments were conducted to investigate the enzymatic characteristics of the purified $\alpha$-amylase (F-2A) of Bacillus circulans F-2 and the digestion rate of various starches. 1. The molecular weight was estimated to be 93000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point was about pH 5.0. The optimum pH for the enzyme action was 6.0-6.5 and the stable pH ranged pH 5.5-12.0. The optimum temperature was 6$0^{\circ}C$, and the purified $\alpha$-amylase was stable below 4$0^{\circ}C$. 2. The purified $\alpha$-amylase was activated by Mn$^{++}$ and Co$^{++}$, whereas it was inhibited by Ag$^{+}$, HT$^{++}$, Cu$^{++}$ and Pb$^{++}$. 3. The purified $\alpha$-amylase is considered to have no sulfhydryl residue essential for its catalytic activity. 4. Michaelis constant (Km) was 1.704 mg/$m\ell$. Activation energy between 25-4$0^{\circ}C$ was 12.297 Kcal/mole, and between 40-6$0^{\circ}C$, it was 7.831 Kcal/mole. 5. The hydrolysis product from soluble starch, amylose and amylopectin in the early stage of hydrolysis was G$_{6}$, and as hydrolysis proceeds, G$_4$and G$_2$appeared. 6. Products from each oligosaccarides are as follows: G$_4$longrightarrow G$_2$+ G$_2$,G$_3$ +G$_1$,G$_{5}$longrightarrow G$_4$+G$_1$,G$_{6}$longrightarrowG$_4$+ G$_2$,G$_{7}$ G$_4$,G$_{8}$longrightarrow G$_4$+G$_4$, 7. On raw potato starch, raw sago starch and raw yam starch, the purified enzyme exhibited a remarkably high digestion rate than Porcine pancreatic amylase and Streptococcus bovis amylase.

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Studies on Protein Profiles and Isozymes in Germinating Seeds (종자발아에 있어서 Protein Profile과 Isozyme에 관한 연구)

  • 권오용
    • Journal of Plant Biology
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    • v.17 no.4
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    • pp.143-156
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    • 1974
  • The purpose of this experiment was to study one side of germination physiology based on that protein profiles and protease relating to protein metabolism, that peroxidase, catalase, $\alpha$-amylase, $\beta$-amylase, and malate dehydrogenase involved in the carbohydrate metabolism of seed germination. All these experiments were divided into the two groups with and without acetone treatment, and were carried out. The protein bands of each germinating stage between the groups treated with and without acetone showed certain basic pattern in polyacrylamide gel disc electrophoresis. However, there was a little difference in the number of protein band, optical density, and migration velocity between two groups. The isozyme bands of peroxidase, and catalase between two groups in polyacrylamide gel disc electrophoresis did not show the numeral difference, but the optical density of certain germinating stage treated with acetone was higher than the group untreated with it and it showed their enzyme activity. The $\alpha$-amylase and $\beta$-amylase activities which involved in starch metabolism of seed germination were higher in the treated group than the other. On one hand, the protease activity of hydrolase occurred in the seeds for germination was also higher, more or less in the treated group than in the other. The isozyme band pattern of malate dehydrogenase in TCA cycle of energy metabolism pathway was very different between two groups growing for 72 hours with and without acetone treatment in cellulose acetate electrophoresis. It indicated that two isozyme bands of malate dehydrogenase was high. Consequently these experimental results mentioned above indicated that acetone treatment before sowing had an effect on dissolving certain complexed lipid substance involved in the seed coats, the activity of carbohydrate hydrolase increased with water absorption which was most comfortable in its germination, dissolved glycerin and fatty acid became certain energy source, and they stimulated the acceleration of respiration metabolism.

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Microbiological Studies on the Rice Makkulli (Part 1) Utilization of Rice Makkulli Koji with the Isolated Strain M-80 (쌀막걸리의 미생물학적 연구 (제1보) 분리균주 M-80의 쌀막걸리 제국용으로서의 이용성)

  • 조용학;성낙계;정덕화;윤한대
    • Microbiology and Biotechnology Letters
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    • v.7 no.4
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    • pp.217-223
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    • 1979
  • Six strains of mold which had high saccharifying and acid-productive ability were isolated from wild sources. The strain M-80 among them and Aspergillus kawachii, which was generally used as rice makkulli koji were used for this studies. The results obtained were summarized as follows. 1) $\alpha$-amylase activities of the strain M-80 and Asp, kawachii showed similarly as about 140W. V., while $\beta$-amylase activity of M-80 was 54 A. U. and Asp. kawachii was 40 A. U. 2) Acid protease activity of M-80 was higher then Asp. kawachii but alkaline protease activity was lower Asp. kawachii respectively. 3) The contents of total acid, ethanol and fusel oil in makkulli brewing with M-80 were higher those of Asp. kawachii and methanol contents of the tested two strains were about 33mg/%. 4) Fifteen kinds of free amino acid were detected from makkulli brewing of two strains, and free amino acid contents of M-80 were 10% higher than those of Asp. kawachii.

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Characterization of the Germinated Rices to Examine an Application Potentials as Functional Rice Processed Foods (기능성 쌀가공식품 원료료의 활용가능성 검토를 위한 발아미의 특성조사)

  • Kang, Mi-Young;Lee, Yeon-Ri;Nam, Seok-Hyun
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.696-701
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    • 2003
  • Rice seeds of 4 cultivars including Whachung and Nampung, of the non-waxy rice cultivars, and Shinsunchalbyeo and Whachungchabyeo, of the waxy rice cultivars, were germinated at $27^{\circ}C$ for 3 days to compare the changes in some physico-chemical properties of the starch granules and the starch-hydrolysing enzyme activities during germination, respectively. With the starch granules, the amount of long glucose chains from amylose molecules were reduced in the non-waxy rices, while the chain length increased in the waxy rices. In the distribution profile of the glucose chain length from amylose molecules, we could observed that the chain length with DP (degree of polymerization) ranged 33 to 66 increased with the decreasing rate of the chain length with that above 130, regardless of the waxiness of rices. In addition, we observed that in contrast to a increase in chain length with DP ranged 14 to 33, the amount of short chains from A chain fraction decreased. Germination induced slight reduction in the polymerization rate of starch granules, and decrease in both initiation and termination temperatures for the gelatinization. ${\alpha}-Amylase$ activity of rices germinated for 3 days found to he higher than that of malt. Especially, the activity of Shinsunchalbyeo was revealed to be highest, about two fold higher than that of malt. In contrast, ${\beta}-amylase$ of the waxy rice found to be considerably less active than malt, although the waxy showed prevalent activity as compared to the non-waxy rices.

A Highly Active Alpha Amylase from Bacillus licheniformis: Directed Evolution, Enzyme Characterization and Structural Analysis

  • Liu, Yihan;Fan, Shuai;Liu, Xiaoguang;Zhang, Zhimeng;Wang, Jianling;Wang, Zhengxiang;Lu, Fuping
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.898-904
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    • 2014
  • The stability of Bacillus licheniformis alpha-amylase (BLA) under acid condition was enhanced through direct evolution using the error-prone polymerase chain reaction. One beneficial mutation site, H281I, was obtained in BLA. The specific activity of H281I was 161/352 U/mg, which was 62.6/27.5% higher than that of the wild-type (WT) (99/276 U/mg) at pH 4.5/6.5 and $95^{\circ}C$. The pH optimum for H281I was decreased about 1 unit, whereas no significant changes of optimum temperature and thermostability were observed compared with the wild type (WT). The $k_{cat}/K_m$ value of H281I was 1.7-/1.4-fold higher at pH 4.5/6.5, respectively, than that of WT. The structure model analysis indicated that the H281I mutation altered the predicted interaction between the amino acid residues at 281 and 273, thus creating a conducive local environment for substrate binding, as reflected by its decreased $K_m$, and consequently increased the specific activity.

Impacts of whey protein on starch digestion in rumen and small intestine of steers

  • Lee, Sang-Bum;Lee, Kyoung-Won;Lee, Jae-Sung;Kim, Kyung-Hoon;Lee, Hong-Gu
    • Journal of Animal Science and Technology
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    • v.61 no.2
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    • pp.98-108
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    • 2019
  • Four Korean native steers ($511{\pm}17.2kg$; $2{\times}2$ replicated crossover design) fitted with duodenal cannulas were used to investigate the influence of oral administration of soluble whey protein (WP; 82.29% crude protein) on ruminal fermentation, gastrointestinal (GI) hormone secretion in the blood, pancreatic ${\alpha}$-amylase activity in the duodenum, and disappearance rate in each segment of the GI tract. Steers were orally fed the basal diet (control; TMR [total mixed ration] 9 kg/d) or the basal diet with enriched WP (400 g/d) for 14 days. The apparent crude protein disappearance rate in the rumen of the WP was higher than in control (p < 0.05). However, no difference between groups was observed in the apparent crude protein disappearance rate in the intestine and the apparent starch disappearance rates in the rumen, GI tract. The level of cholecystokinin, secretin, and ghrelin in serum and pancreatic ${\alpha}$-amylase activity in the duodenum of the WP also did not change. The changes in the level of blood urea nitrogen related to protein metabolism were higher in the WP than in the control (p < 0.05). However, the levels of total protein, lipid, carbohydrate and mineral metabolites did not change. Consequently, we suggest that the oral administration of WP in steers assisted in ruminal fermentation due to the population increase of microbes in the rumen but did not improve the starch digestion rate in the small intestine because GI hormone secretion in the blood and pancreatic ${\alpha}$-amylase activity did not change.

Studies on Antibiotic Producers of Korean Soil Microbes(I) -Isolation and Antibiotic Activity of Streptomyces Strain DMC-72- (한국(韓國) 토양균(土壤菌)중 항생물질(抗生物質) 생성균(生成菌)에 관한 연구(硏究)(제1보)(第1報) -스트렙토마이세스속(屬) 균주(菌株) DMC-72호(號)의 분리(分離) 및 항균작용(抗菌作用)-)

  • Kim, Kwang-Wook;Choi, Eung-Chil;Shim, Mi-Ja;Kim, Byong-Kak
    • Korean Journal of Pharmacognosy
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    • v.15 no.1
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    • pp.15-23
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    • 1984
  • To find antimicrobial strains of the soil microorganisms in Korea, they were isolated from the soil samples of different locations and screened for antibiotic activity against several standard microbes. An isolate among them had an antibacterial activity against gram-positive bacteria. The examination of its morphological and biochemical characteristics according to the International Streptomyces Project methods showed that it belongs to the genus Streptomyces. The strain was named DMC-72. The strain appears to be a new strain when it was compared with the species within the genus which have been so far reported. The antibiotic metabolite of the strain was produced in submerged culture method. It was found to be a quinone compound and was named soulomycin. This strain was also found to produce an ${\alpha}-amylase$ inhibitor in the submerged culture.

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In vitro safety and efficacy of probiotics mixture on carbohydrate digestion inhibition (프로바이오틱스 혼합물의 in vitro에서의 안전성 및 탄수화물 소화 저해 효능 평가)

  • Eunsol Seo;Jang-Bin Woo;MinYeong Seo;Jeongmin Woo
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.538-545
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    • 2023
  • This study aimed to assess the in vitro safety of a probiotics mixture (Lactobacillus acidophilus PBS066, Lactiplantibacillus plantarum PBS067, and Limosilactobacillus reuteri PBS072), along with its inhibitory effect on carbohydrate digestion. All three strains met the antibiotic resistance profile of the European Food Safety Authority (EFSA) guidelines. None of the strains exhibited hemolytic activity or cytotoxicity against Caco-2 cells. Strains PBS067 and PBS072 inhibited α-amylase activity, whereas all three strains suppressed α-glucosidase activity, indicating that the mixture might limit carbohydrate digestion in the gastrointestinal tract. These findings support the safety of this probiotics mixture and its potential to modulate carbohydrate metabolism in the gut.

Antioxidative and anti-α-amylase activities of four wild plants consumed by pastoral nomads in Egypt

  • Hossain, SJ;El-Sayed, M;Aoshima, H
    • Advances in Traditional Medicine
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    • v.9 no.3
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    • pp.217-224
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    • 2009
  • In this study, four plants grown naturally in Egypt that are usually consumed by pastoral nomads were screened for their total phenolic contents, antioxidative, and anti-$\alpha$-amylase activities. Dried powder of plant's part was extracted in absolute or 70% ethanol. A polar extract of Panicum turgidum (PTPE) had the highest total polyphenol content {92.5 mg gallic acid equivalents (GAE)/g}, followed by an alkaloid extract of Withania somnifera (WSAlk; 77.5 mg GAE/g), and an ethanol extract of Leptadenia pyrotechnica (LPEE; 59.1 mg GAE/g). By employing different assays such as DPPH radical scavenging, reducing power, $Fe^+$ chelating, $H_2O_2$ scavenging and total antioxidant capacity, it was shown that PTPE, WSEE (ethanol extract of W. somnifera), WSAlk and LPEE had promising antioxidant activity, though, their potency varied according to the different tests. WSAlk had the highest level of $\alpha$-amylase inhibition (40.2%) in vitro, followed by WSEE (30.5%). Therefore, it can be concluded that these plants, especially extracts of PTPE, and WSAlk are beneficial to physiological health, and could be used in food and pharmaceutical industries to prepare dietary supplements, functional foods or food preservatives.