• Title/Summary/Keyword: ${\alpha}$-amylase activity

Search Result 563, Processing Time 0.022 seconds

Cloning and Expression of an $\alpha$-Amylase Gene from Bacillus circulans in B. subtilis and B. megaterium (Bacillus circulans $\alpha$-amylase 유전자의 Basillus subtilis와 Bacillus megaterium에서의 클로닝 및 발현)

  • 이동석;김지연;김한복
    • Korean Journal of Microbiology
    • /
    • v.36 no.3
    • /
    • pp.203-208
    • /
    • 2000
  • A Baczllus circdans KCTC3004 $\alpha$-amylase gene contained in a recombinant plasmid pAL850 was transferred into a new shuttle vector plasmid pALSIlI by ligating linearlzed DNAs of pUC19 and pUB110. B. subtilis RM125 and B. megatenurn ATCC14945 transfonned with pALS111 produced the $\alpha$-amylase substantially Most of the enzyme was produced during the exponential growth period. The maxiinurn activities of the $\alpha$-amylase produced by the Bucillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125(pALS111) enzyme showed the actlvicy 95 times higher than that of the gene donor cells, followed by the B, nzegaterium ATCC14945(pALSlll) enzyme with activity 34 limes higher than that of the gene donor cells. While E coli secreted about 10% of the produced enzyme, B. subtilis excreted the enzyme inlo the medium wholly and B. megaterirun about 98% ofthe total product. The plasmid pALSI11 was quite stable inB. nzegaterium (92%), inoderately stable in B. subtilis (76%), but was unstable in E. coli (38%). The SDS-PAGE and zymogram of this enzyme produced in E. coli(pALS111), B. subtilis( pALS111) or B. megateril~m (pALS111) indicated a molecular weight of 55,000. The enzymes overproduced in three different host cells hydrolyzed starch to produce mainly maltoaiose and mallooligosaccharides.

  • PDF

Antioxidant and Anti-diabetes Activities of Methanolic Extract and Fractions of Astragalus membranaceus Roots

  • Park, Jae-Hyo;Yin, Yu;Wang, Myeong-Hyeon
    • Preventive Nutrition and Food Science
    • /
    • v.15 no.1
    • /
    • pp.30-35
    • /
    • 2010
  • The potential biological activities of methanol extract and 5 fractions (hexane, $CH_2Cl_2$, EtOAc, BuOH and water) from roots of Astragalus membranaceus were examined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, hydroxyl radical (${\cdot}OH$) scavenging activity, reducing power assays, lipid peroxidation inhibitory activity, $\alpha$-amylase and $\alpha$-glucosidase inhibition assays. The EtOAc fraction showed high DPPH free radical scavenging activity ($EC_{50}=170.34\;{\mu}g/mL$), hydroxyl radical scavenging activity ($EC_{50}=32.14\;{\mu}g/mL$), lipid peroxidation inhibitory activity ($EC_{50}=52.46\;{\mu}g/mL$) and a concentration dependence, with OD value ranging from 0.234 to 0.345 (0.1 to 0.5 mg/mL), for reducing power. The EtOAc fraction has the highest total phenolic content ($142.13\;Gal\;{\mu}g/mg$) and the $CH_2Cl_2$ fraction has the highest flavonoid content ($71.63\;Que\;{\mu}g/mg$). Meanwhile, hexane and EtOAc showed certain $\alpha$-amylase and $\alpha$-glucosidase inhibition activities. These results suggest that the methanol extract and fractions from Astragalus membranaceus root have significant antioxidant and anti-diabetes activities, which could be used as a potential source of pharmaceutical materials.

Antidiabetes and Angiotensin Converting Enzyme Inhibitory Activity of Sonchus asper (L) Hill Extract (큰방가지똥 추출물의 항당뇨 및 항고혈압효과)

  • Xu, Ming-Lu;Wang, Lan;Xu, Gui-Fang;Wang, Myeong-Hyeon
    • Korean Journal of Pharmacognosy
    • /
    • v.42 no.1
    • /
    • pp.61-67
    • /
    • 2011
  • In this study, we evaluated the bioactivities of methanol extract and its solvent fractions of Sonchus asper (L.) Hill. The EtOAc fraction of S. asper exhibited more strong antioxidant activity than other extracts as evidenced by the strongest 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity with a $EC_{50}$ value at $33.55\;{\mu}g/mL$ and reducing power, the total polyphenol (180.71 mg GAE/g) and flavonoid contents (145.86 mg QE/g) of S. asper extract were higher than other extracts. The EtOAc fraction of the S. asper also showed 47.38% mushroom tyrosinase inhibition activity, 56.22% ${\alpha}$-glucosidase inhibition and 46.58% ${\alpha}$-amylase inhibition ratio at 1 mg/mL. Both methylene chloride and EtOAc fractions of methanol extract of S. asper effectively reduced of the 86.34% and 62.03% angiotensin I converting enzyme (ACE) activity at 2 mg/mL, respectively. These findings suggest that the EtOAc fraction of the S. asper could be a potential antioxidant in food additive, medicinal, and industry product.

Stabilization of Barley $\beta$-Amylase by Modification with $IO_4$-Oxidized Starch ($IO_4$-산화전분 변형에 의한 보리 $\beta$-Amylase의 안정성 증가)

  • 안용근
    • The Korean Journal of Food And Nutrition
    • /
    • v.13 no.4
    • /
    • pp.342-347
    • /
    • 2000
  • The stabilization of barley $\beta$-amylase(Biozyme ML, Amano) was attained by modification with periodate-oxidized soluble starch. The specific activities of modified enzyme at pH 9.7 and pH 8.0 were 42% and 92%, respectively, compared with that of native enzyme. The pH stability of modified enzyme was increased at pH 2~5 and 7~12 in the presence of $\alpha$-cyclodextrin( $\alpha$ -CD) compared wish that of native enzyme. Thermal stability of the modified enzyme was increased. After treatment at 6$0^{\circ}C$ for 10min. the activity remained 8% for the enzyme modified at pH 8.0 in the presence of $\alpha$-CD, 4.5% for the native enzyme. The native enzyme and modified enzyme showed two peak in HPLC. The molecular weight of the modified enzyme was slightly increased in HPLC analysis.

  • PDF

Proximate Compositions and Biological Activities of Lactuca indica L. Seonhyang and Wild Species Depending on Harvesting Time (왕고들빼기 선향과 야생종의 수확시기별 일반성분 및 생리활성)

  • Kwon, Hye-Jeong;Jeong, Eun-Kyoung;Jeong, Hye-Jeong;Kim, Si-Chang;Heo, Nam-Kee;No, Hee-Sun
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.44 no.3
    • /
    • pp.363-369
    • /
    • 2015
  • We investigated the effect of harvesting time on the proximate compositions and biological activities of Lactuca indica L. Seonhyang and wild species. The highest moisture content (91%) was obtained from Seonhyang harvested in June. The wild species harvested in August showed the highest crude protein (4.8%) and crude fiber contents (2.7%). Potassium contents were 626~684 mg/100 g, and no significant difference in harvesting time was observed between Seonhyang and wild species. Calcium and phosphate contents of Seonhyang leaves harvested in August were highest (350 mg/100 g and 123 mg/100 g, respectively). The highest total polyphenol and total flavonoid contents were observed for wild species harvested in June (60 mg/g and 126 mg/g, respectively). Ethanol extracts of Seonhyang and wild species leaves harvested in June showed the highest DPPH radical scavenging activity (95%). ${\alpha}$-Amylase inhibitory activity was highest (94.8%) in water extract of Seonhyang leaves harvested in July. Nitric oxide production inhibitory activity was $14.3{\mu}M$ in ethanol extracts of Seonhyang and $16.8{\mu}M$ in ethanol extracts of wild species harvested in June. Calcium content and ${\alpha}$-amylase inhibitory activity of Seonhyang leaves were greater than those of wild species leaves. These results suggest that Seonhyang leaf can be used to develop processed foods.

$\alpha$- and $\beta$-Amylase Isozyme Expresser Native Proteins in Tropical Silkworm Bombyx mori L.

  • Chattopadhyay, G.K.;Verma, A.K.;Sengupta, A.K.;Das, S.K.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.8 no.2
    • /
    • pp.189-194
    • /
    • 2004
  • Amylase isozyme based three multivoltine viz., N+p, Np, N+ $p^{cho}$ and two bivoltine-D6+p, D6p syngenic lines (Syn. L) were developed from germplasm (GP) stocks Nistari (N) and D6 respectively. haemolymph isozyme pattern at pH 7.0 and 8.5 depicted a total 11 number (Am $y_{1 to 6}$ at pH 7.0 and Am $y^{l to 5}$ at pH 8.5) of native proteins (NP) of various sizes are amylase isozyme expressers. Among eleven NPs, two NPs of 770 kDa (Am $y^{6}$ at pH 7.0) and 376 kDa (Am $y^3$ at pH 8.5) are $\alpha$-amylase expressers and remaining NPs of 370, 364, 350, 329 and 274 kDa at pH 7.0 and 206, 292, 416, 725 kDa at pH 8.5 are $\beta$-amylase expressers. Accordingly, digestive juice amylase isozyme pattern at aforesaid pH also depicted a total number of 10 NPs (Am $y^{1 to 5}$) at each pH 7.0 and 8.5 are amylase expressers of which NP of 387 kDa (Am $y^4$ at pH 7.0) and 780 kDa (Am $y^{5}$ at pH 8.5) are a-amylase expresser. Remaining NPs of 338,297 & 216 kDa at pH 7.0 and 370, 341, 329 &302 kDa at pH 8.5 are $\beta$-amylase expresser. Recurrent backcross lines (RBL) viz., N+pRBL and NpRBL were developed through introgression of high shell weight character (a multigenic trait) to be used further for congenic line (Con. L) development and to understand any association with introgressed character. Isozyme pattern in haemolymph of RBLs depicted only one $\alpha$-amylase of 770 kDa at pH 7.0 and 376 kDa at pH 8.0 with three and four respective $\beta$-amylase bands but in bivoltine lines numbers of $\beta$-amylase bands vary between 1 to 2 at aforesaid pH. Variability was also observed in digestive juice of multivolitine and its RBLs but bivoltine lines express null activity at both pH except appearance of one very week $\alpha$-amylase band D6+p at pH 8.5. Overall study suggests that not a single NP at both pH is common for expression of any band of amylase isozyme i.e., a totally different set of proteins are the amylase isozyme expresser at specific pH and no molecular factor of amylase is associated in developed RBLs which showed improvement on survival, single cocoon shell weight (SCSW) and single filament length over receptor parents.s.s.s.

Isolation of Mutants Overproducing Amylase from Nuruk Fungi by NTG (NTG에 의한 Amylase활성이 높은 누룩사상균의 변이주의 분리)

  • 정혁준;김영숙;유대식
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.29 no.6
    • /
    • pp.987-994
    • /
    • 2000
  • Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg. ̄) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade. ̄) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.

  • PDF

Antioxidative activity, including Inhibitory activities of ACE, APN and $\alpha$-amylase, in Theaceae Plants Native to Jeju Island (제주도 자생 차나무과 식물의 ACE, APN, $\alpha$-amylase 저해 활성 및 항산화활성에 대한 연구)

  • Oh, Soon-Ja;Lee, Jin-Ho;Ko, Kwang-Sup;Shin, Dong-Bum;Koh, Seok-Chan
    • Korean Journal of Plant Resources
    • /
    • v.23 no.5
    • /
    • pp.406-414
    • /
    • 2010
  • Antioxidative activity, including inhibitory activities of angiotensin I converting enzyme(ACE), aminopeptidase N(APN) and $\alpha$-amylase, was investigated in the methanol extracts from Theaceae plants native to Jeju island, in order to select the plant species containing bioactive materials for functional food or medicines. ACE inhibitory activity was above 50% in Ternstroemia japonica(stem bark) and Cleyera japonica(leaf), and APN inhibitory activity was low to be positive only in C. japonica(leaf, stem bark) and T. japonica(stem bark). $\alpha$-Amylase inhibitory activity was above 30% in Camellia japonica(fruit), Eurya emarginata(stem), T. japonica(stem bark) and Thea sinensis(stem). The antioxidative activity, estimated by the DPPH radical scavenging capacity, was above 30% in C. japonica(stem bark), T. japonica(stem bark) and T. sinensis(leaf). Particularly, the antioxidative activity analyzed by dot-blot test was very high in C. japonica(stem bark) relatively to those of other plants, and remained high in the low concentration($1.25\;{\mu}g/m{\ell}$). From the TLC analysis of antioxidative compounds, EGC(Rf 0.26) was found to have high activity in stem bark of C. japonica and EGCG(Rf 0.09) was found to have high activity in stem bark of C. japonica, E. emarginata, and T. japonica. Five bands (Rf 0.54, 0.46,0.44, 0.16, 0.03) which were not identified as compared with catechins were detected as polyphenolic compounds on the TLC plates sprayed with the Folin-Ciocalteu solution or the Ferric chloride-alcohol solution. These results suggests that Theaceae plants except E. japonica could be potentially used as a resource of bioactive materials for functional foods or medicines and further research is reguired to identify the bioactive substances and determine the functions of them.

Effect of Oral Administration of Intact Casein on Gastrointestinal Hormone Secretion and Pancreatic α-Amylase Activity in Korean Native Steer

  • Lee, S.B.;Choi, C.W.;Jin, Y.C.;Wang, T.;Lee, K.H.;Ku, M.B.;Hwang, J.H.;Kim, K.H.;Vega, R.S.A.;Lee, H.G.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.26 no.5
    • /
    • pp.654-660
    • /
    • 2013
  • Three Korean native steers ($779{\pm}24$ kg) fitted with duodenal cannulas were used in a $3{\times}3$ Latin square design to investigate the influence of oral administration of soluble proteins, intact casein (IC) and acid hydrolyzed casein (AHC), on gastro-intestinal hormone (GIH) secretion in the blood and pancreatic ${\alpha}$-amylase activity in the duodenum. Oral treatment consisted of a basic diet (control), IC (C+100% protein), or AHC (C+80% amino acid, 20% peptide) for 21 d. Blood and duodenum samples were collected for measurement of serum GI hormones, and pancreatic ${\alpha}$-amylase activity was determined at 900, 1030, 1330, 1630, and 1930 h after feeding on d 21 of treatment. The levels of serum cholecystokinin (CCK) and secretin in the IC treatment group were higher compared to the other treatment groups (p<0.05). In addition to the changes in CCK and secretin levels upon IC treatment, the pancreatic ${\alpha}$-amylase activity in the duodenum was higher in the IC group compared to the control diet group (p<0.05). The response of serum ghrelin to IC and AHC treatment was in accordance with the response of serum secretin. The level of peptide fragments flowing in the duodenum was higher in the IC treatment group than the other treatment groups (p<0.05). In conclusion, this study demonstrated that an increase in duodenal CCK and secretin upon IC oral administration increased pancreatic ${\alpha}$-amylase secretion. In addition, ghrelin may be associated with GI hormone secretion in Korean native steers.

Changes in Microflora, Enzyme Activities and Microscopic Structure of Waxy Rice and Steeping Water in Response to Different Steeping Conditions During Preparation of Gangjung (강정 제조시 찹쌀의 수침조건에 따른 찹쌀과 수침액의 미생물상, 효소활성 및 미세구조 변화)

  • Kim, Haeng-Ran;Kim, Kyung-Mi;Kim, Kwang-Ok
    • Korean Journal of Food Science and Technology
    • /
    • v.41 no.6
    • /
    • pp.644-651
    • /
    • 2009
  • This study was conducted to investigate the effects of steeping periods and temperatures of waxy rice on the microflora and enzyme activities of steeping water and waxy rice, as well as on the microscopic structures of waxy rice during the preparation of gangjung. When the steeping water was 15 and $35^{\circ}C$, yeast and lactic acid bacteria were the major microflora after 1 day. After 11 days, Lactobacillus spp. were the major bacteria, while Bacillus spp. were the primary microflora after 21 days. At $25^{\circ}C$ the primary species were Bacillus spp., Leuconostoc spp., and Bacillus spp. after 1, 11 and 21 days, respectively. The $\alpha$-amylase activity of steeping water increased proportionally with the increase of temperature and period, while the protease activity tended to increase for 11 days, but decreased at 21 days. The enzyme activities of the steeped waxy rice, especially the $\alpha$-amylase activity, decreased with higher steeping temperature, while the $\beta$-amylase and glucoamylase activity remained constant for up to 21 days. Observation using scanning electron microscopy indicated that intracellular cleavage of steeped waxy rice endosperm resulted in a rough surface morphology due to the exposure of polygonal starch granules. Tiny pinholes on the surface of waxy rice starch granules were also observed after steeping for 11 days.