• 제목/요약/키워드: fungal a-amylase

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내산성(耐酸性) 소화효소제(消化酵素劑)의 생산(生産)에 관(關)한 연구(硏究) -내산성(耐酸性) 효소생산균(酵素生産菌)의 분리(分離)와 효소(酵素) 생산조건(生産條件)에 관(關)하여- (Studies on the Production of Acid Digestive Enzyme -Isolation and Characterization of a Fungal Strain Which Produces Acid Enzymes-)

  • 손천배;박윤중
    • 한국식품과학회지
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    • 제13권3호
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    • pp.241-246
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    • 1981
  • 소화효소제(消化酵素劑) 생산(生産)에 이용(利用)하기 위하여 공기(空器)중(中)에서 내산성(耐酸性) protease 및 amylase 생산능(生産能)이 강(强)한 균주(菌株)를 분리(分離), 선정(選定)하고 선정균(選定菌)의 균학적(菌學的) 성질(性質)을 검토(檢討)하였다. 아울러 생성효소(生成酵素)의 내산성(耐酸性)(반응(反應) pH와 효소활성(酵素活性)과의 관계(關係)과 효소(酵素) 생산조건(生産條件)을 검토(檢討)하였다. 1. 선정균주(選定菌株)는 내산성(耐酸性) protease 및 amylase를 강(强)하게 생산(生産)하는 균주(菌株)로서 Raper와 Fennel의 manual에 의(依)하여 Aspergillus niger로 동정(同定)되었다. 2. 선정균(選定菌)의 protease는 pH 2.0에서 최대활성(最大活性)을 나타냈으며, ${\alpha}-amylase$$pH4{\sim}5$, glucoamylase는 $pH3{\sim}5$에서 최대활성(最大活性)을 나타냈다. 3. 밀기울 배지(培地)에 배양시(培養時) protease(pH 2.5에서의 활성)생산(生産)의 최적조건(最適條件)은 $30^{\circ}C$, $2{\sim}3$일간(日間)이며 , ${\alpha}-amylase$ 및 glucoamylase(pH3.0 에서의 활성(活性))의 경우는 $30^{\circ}C$, 3일간(日間)이었다. 4. 밀기을배지(培地)에 옥수수전분을 2% 첨가(添加)한 경우 내산성(耐酸性) protease 및 glucoamylase의 생성(生成)이 약(約) 20%씩 증가(增加)되었다. 5. 밀기울배지(培地)에 황산암모늄 0.3%를 첨가(添加)한 경우 내산성(耐酸性) protease 및 glucoamylase의 생성(生成)이 증가(增加)되었으며, 특(特)히 내산성(耐酸性) protease의 생성(生成)에 효과적(效果的)이었다.

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Characterization of Alpha Amylase Producing Thielaviopsis ethacetica and Its Raw Starch Hydrolyzing Ability on Different Agricultural Substrates

  • Dissanayaka, Dissanayaka M.S.;De Silva, Sembukuttige N.T.;Attanayaka, D.P.S.T.G.;Kurera, Mihidukulasuriya J.M.S.;Fernando, Charakrawarthige A.N.
    • 한국미생물·생명공학회지
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    • 제47권3호
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    • pp.412-422
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    • 2019
  • The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae

  • Yin, Yanchen;Mao, Youzhi;Yin, Xiaolie;Gao, Bei;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • 제25권7호
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    • pp.988-998
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    • 2015
  • The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

곰팡이 균종을 달리하여 제조한 혼합 곡류 누룩의 품질특성 (Assessment of the Quality Characteristics of Mixed-grain Nuruk Made with Different Fungal Strains)

  • 백성열;김주연;최지호;최정실;최한석;정석태;여수환
    • 동아시아식생활학회지
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    • 제22권1호
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    • pp.103-108
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    • 2012
  • 밀, 쌀 및 녹두를 분쇄 후 밀기울을 첨가하여 일정 비율(15:1:1:3)로 혼합한 곡물에 양조용 곰팡이의 균종을 달리하여 제조한 누룩의 품질 특성을 알아보고자 발효 시기별 일반성분 및 효소활성과 유기산 분석을 하였다. 누룩의 pH는 발효 시기별 큰 차이는 보이지 않았으며, 산도와 아미노산도는 두균을 혼합하여 제조한 누룩(AO-AK)이 가장 높게 나타났다. 전반적으로 두 균을 혼합한 누룩이 단일 균을 접종한 누룩보다 ${\alpha}$-amylase 및 acidic protease의 효소활성이 높게 나타났다. 누룩의 유기산은 acetic, citric, formic, fumaric, lactic, malic 및 oxalic acid 등이 확인되었다. 전체 유기산 총량은 혼용누룩(2,116.3 mg%), A. kawachii SC60 단용누룩(1,608.5 mg%), A. oryzae RIB1353 단용누룩(1,146.7 mg%) 순으로 혼용누룩의 유기산 생성량이 가장 높게 나타났다. 따라서 두 균주를 혼용하여 제조한 누룩이 단일균주를 사용한 누룩보다 유기산 함량, 효소활성 등 품질 특성이 우수한 것으로 여겨진다.

Enzyme Activities and Substrate Degradation by Fungal Isolates on Cassava Waste During Solid State Fermentation

  • Pothiraj, C.;Eyini, M.
    • Mycobiology
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    • 제35권4호
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    • pp.196-204
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    • 2007
  • The growth and bioconversion potential of selected strains growing on cassava waste substrate during solid state fermentation were assessed. Rhizopus stolonifer showed the highest and the fastest utilization of starch and cellulose in the cassava waste substrate. It showed 70% starch utilization and 81% cellulose utilization within eight days. The release of reducing sugars indicating the substrate saccharification or degradation potential of the organisms reached the highest value of 406.5 mg/g by R. stolonifer on cassava waste during the eighth day of fermentation. The protein content was gradually increased (89.4 mg/g) on the eighth day of fermentation in cassava waste by R. stolonifer. The cellulase and amylase activity is higher in R. stolonifer than A. niger and P. chrysosporium. The molecular mass of purified amylase and cellulase seemed to be 75 KDal, 85 KDal respectively.

Structural Investigation and Homology Modeling Studies of Native and Truncated Forms of $\alpha$-Amylases from Sclerotinia sclerotiorum

  • Ben Abdelmalek, Imen;Urdaci, Maria Camino;Ali, Mamdouh Ben;Denayrolles, Muriel;Chaignepain, Stephane;Limam, Ferid;Bejar, Samir;Marzouki, Mohamed Nejib
    • Journal of Microbiology and Biotechnology
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    • 제19권11호
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    • pp.1306-1318
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    • 2009
  • The filamentous ascomycete Sclerotinia sclerotiorum is well known for its ability to produce a large variety of hydrolytic enzymes. Two $\alpha$-amylases ScAmy54 and ScAmy43 predicted to play an important role in starch degradation were showed to produce specific oligosaccharides essentially maltotriose that have a considerable commercial interest. Primary structure of the two enzymes was established by N-terminal sequencing, MALDI-TOF masse spectrometry and cDNA cloning. The two proteins have the same N-terminal catalytic domain and ScAmy43 derived from ScAmy54 by truncation of 96 amino acids at the carboxyl-terminal region. Data of genomic analysis suggested that the two enzymes originated from the same $\alpha$-amylase gene and that truncation of ScAmy54 to ScAmy43 occurred probably during S. sclerotiorum cultivation. The structural gene of Scamy54 consisted of 9 exons and 8 introns, containing a single 1,500-bp open reading frame encoding 499 amino acids including a signal peptide of 21 residues. ScAmy54 exhibited high amino acid homology with other liquefying fungal $\alpha$-amylases essentially in the four conserved regions and in the putative catalytic triad. A 3D structure model of ScAmy54 and ScAmy43 was built using the 3-D structure of 2guy from A. niger as template. ScAmy54 is composed by three domains A, B, and C, including the well-known $(\beta/\alpha)_8$ barrel motif in domain A, have a typical structure of $\alpha$-amylase family, whereas ScAmy43 contained only tow domains A and B is the first fungal $\alpha$-amylase described until now with the smallest catalytic domain.

Fungal bioconversion of Korean food wastes for the production of animal feed additive enzymes

  • 정윤승;정상원;조아라;권순우;한승호
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.529-532
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    • 2001
  • Korean food waste, one of the abundantly available but environmentally problematic organic wastes in Korea, was utilized as solid-substrate by fungal strain Aspergillus niger ATcC 6275 for the production of enzymemixture containing amylase, cellulase and xylanase. The enzyme mixture can be used as high value-added animal feed. Solid-state fermentation method yielded a 84-fold enhancement in xylanase activity compared with submerged fermentation method. The effect of incubation period, incubation temperature, pH of medium, moisture content, inoculum size and enrichment of the medium with nitrogen and carbon sources were observed for optimal production of these enzymes The optimal amylase activity of 33.10 U/g, cellulase activity of 24.41 U/g, xylanase activity of 328.84 U/g were obtained at 8 days incubation with 50%(w/w) soy bean flake, with incubation temperature of $25^{\circ}C$, pH of 6.38, optimal moisture content of 55% and with inoculum size of $3.8{\times}10^6$spore/g. Enzyme activities were enhanced when ImM $CaSO_4$, 2% Malt extract and 2% galactose were added as mineral, nitrogen and carbon enrichment respectively.

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Extrusion시킨 팽윤 전분을 기질로 한 새로운 Maltose 생산법 (A Novel Method for Production of Concentrated Purity Maltose Using Swollen Extruded Starch)

  • 이용현;김동선;신현동;박진서
    • 한국미생물·생명공학회지
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    • 제22권1호
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    • pp.106-113
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    • 1994
  • A novel method for production of concentrated purity maltose using swollen extruded corn starch was investigated. Degree of gelatinization of extruded starch suitable for maltose formation was found to be around 70%. The optimal amiunt of enzyme was 400 unit fungal $\alpha $-amylase per g of starch, and the reaction time was 12 hours. At extruded starch concentration of 300 g/l(w/v), maltose concentration and content were reached up to 220 g/l(w/v) and 77%(w/w), respectively. The maltose forming reaction was also successfully proceeded at high starch concentration of 700 g/l(w/v), however, the conversion yield and content were decreased. By the addition of extruded starch by fed-batch wise, the maltose concentration, purity, and conversion yield could be improved up to 465 g/l(w/v), 70%(w/w), and 0.63, respectively. The investigated maltose production process seems to have many potential advantages over the conventional process utilizing liquefied starch, and the feasibility for industrial application needs to be evaluated.

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순창군 장류로부터 분리된 황국균의 동정 및 특성 (Identification and Characterization of Aspergillus oryzae Isolated from Soybean Products in Sunchang County)

  • 임은미;이지영;모하메드;한갑훈;이보순;조용식;김현영
    • 한국균학회지
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    • 제42권4호
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    • pp.282-288
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    • 2014
  • 본 연구에서는 순창지역에서 만들어지는 장류에서 곰팡이를 분리하고 동정하여 보다 안전하고 기능성이 높은 발효제품을 위한 균주를 확보하고자 하였다. 순창지역에서 생산되는 장류 제품으로부터 곰팡이를 분리하여 ${\beta}$-tubulin 유전자 분석 통해 10개의 균주가 Aspergillus oryzae/flavus complex임을 알 수 있었다. 보다 정확한 동정을 위하여 아플라톡신 클러스터 유전자 중에 하나인 omtA의 염기서열을 증폭하여 A. oryzae와 A. flavus 표준 균주의 omtA 서열과 함께 계통 분류한 결과, A. oryzae의 표준 균주와의 유연관계가 높음을 알 수 있었다. 또한 norB-cypA 사이의 염기서열을 증폭한 결과 500 bp이 증폭 산물이 확인되었는데 이는 표준 균주인 A. oryzae의 norB-cypA 사이의 염기서열 증폭 산물과 동일한 크기임을 확인할 수 있었다. A. oryzae로 확인된 10균주를 활용하여 코지를 제조하고 ${\alpha}$-amylase 활성과 protease 활성을 측정하였다. Protease 활성은 6, 13, 17, 27, 37, 그리고 38 균주로 제조된 코지는 대조구(시판되고 있는 종균으로 제작한 코지)보다 2배 정도 높은 protease 활성을 보였으며, ${\alpha}$-amylase 활성은 257~320 U/mL로 측정되었다. 식품안전성을 위한 아플라톡신 분비 확인 결과, 63번 균주로 제조된 코지를 제외한 모든 코지에서 아플라톡신을 만들지 않는 것으로 확인되어, 순창에서 분리된 A. oryzae는 추후 메주 접종균으로 개발할 수 있음을 보여주었다.

Effects of dietary supplementation with different levels and molecular weights of fungal β-glucan on performances, health and meat quality in broilers

  • Kovitvadhi, Attawit;Chundang, Pipatpong;Tirawattanawanich, Chanin;Prathumpai, Wai;Methacanon, Pawadee;Chokpipatpol, Krith
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권10호
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    • pp.1548-1557
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    • 2019
  • Objective: To investigate the effects of dietary supplementation with different levels and molecular weights of fungal ${\beta}$-glucan on productive performances, health, carcass traits and meat quality in broilers. Methods: Two hundred and ten of one-day-old chicks with equal sex were assigned to seven experimental groups in $2{\times}4$ factorial arrangement. These groups were supplemented with (0, 10, 30, and 60 ppm) of molecular weight 1-3, 1-6 ${\beta}$-glucan (low or high). High molecular weight ${\beta}$-glucan (H: 943 kDa) was obtained from Ophiocordyceps dipterigena BCC 2073, whereas H with ${\gamma}$-Irradiation treatment was performed to achieve low molecular weight ${\beta}$-glucan (L: 8 kDa). Results: There was no statistical significance in productive performances, apparent digestibility and interaction between fixed factors along 42 days of experiment (p>0.05). A higher caecal amylase activity was present in the group that received L, while there was a dramatic decrease in H and the control groups, respectively (p<0.05). The increase of supplemental dose increased caecal amylase activity (p<0.05). Immunomodulatory effects from L was revealed by the marked increase of phagocytic activity, relative weight of thymus and bursa of fabricius (p<0.05). Similarly, the additive dose at 30 ppm provided the same results, whereas the only significant difference with supplementation at 60 ppm was an increase in phagocytic activity (p<0.05). Interestingly, villi height of broilers fed L was higher than other groups (p<0.05). The treatments did not influence haematology, blood chemistry, antibody production level against vaccination, carcass traits and meat quality (p>0.05). Conclusion: The supplementation of L at 30 ppm was suggested to achieve benefits of immune modulation without adverse effects on other parameters.