• Title, Summary, Keyword: Aspergillus ficuum

Search Result 18, Processing Time 0.031 seconds

Purification of Phytase from Aspergillus ficuum and Production of Anti-phytase Antibody (Aspergillus ficuum의 Phytase의 정제와 Anti-phytase 항체생산)

  • Kim, Keun
    • The Korean Journal of Mycology
    • /
    • v.27 no.4
    • /
    • pp.299-303
    • /
    • 1999
  • Phytase(myo-inositol-hexakis phosphate 3-phosphohydrolase, E C 3.1.3.8) sequentially hydrolyzes phytate to myo-inositol and inorganic phosphate. Phytase of Aspergillus ficuum was purified to homogeneity using ultrafiltration, cation exchange column and anion exchange column. It's molecular weight is estimated as around 90,000 by SDS-PAGE. Antibody against the phytase was produced by immunizing mice with the purified phytase. The titer of the antibody was determined to be 1/25,000.

  • PDF

Purification and Properties of Aspergillus 3cuum exoinulinase (Aspergillus ficuum 조효소액으로부터 Exoinulinase의 정제 및 특성)

  • 한상배;송근섭;유향숙;노민환;이태규;손희숙;우순자;엄태봉
    • Microbiology and Biotechnology Letters
    • /
    • v.19 no.3
    • /
    • pp.253-258
    • /
    • 1991
  • - An exoinulinase (EC 3.2.1.80) was purified from a commercial inulinase preparation from Aspergillus ficuum using ion exchange chromatography on CM-Sephadex C-50 and DEAESepharose 6B and HPLC gel filtration on a Protein Pak 125 column. Native exoinulinase had a molecular weight of 83, 000$\pm$ 1, 000 and was glycoprotein. Optimal pHs of the enzyme were ranged from 4.4 to 4.7. About ninety five percent of the whole activity was maintained even after incubation of 8 hours at $55^{\circ}C$.The enzyme was a typical non-specific P-fructofuranosidase, of which I/S ratio appears to be 0.35.

  • PDF

Screening of Phytase Overproducing Strains in Aspergillus spp. by UV Mutagenesis

  • Lee, Eung-Suek;Paik, In-Kee;Hahm, Young-Tae
    • Mycobiology
    • /
    • v.28 no.3
    • /
    • pp.119-122
    • /
    • 2000
  • Phytases (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) are enzymes which catalyze the hydrolisys of phytate into myo-inositol and inorganic phosphates. Phytases are found in plants and a variety of microorganisms. Aspergillus species were treated with 254 nm of UV irradiation for the screening of phytase overproducing mutant strains. At 15 minute irradiation, the survivals of population were less than 5%, and UV irradiation time was decided at 20 minute for the isolation of mutant strains. Four UV mutant strains in A. oryzae (YUV-47, -169, -341, -511) and six in A. ficuum (FUV-17, -36, -69, -193, -317, -419) were isolated on PSM media containing ammonium phosphate. The specific enzyme activities of A. ficuum mutants are 110 to 140% higher than that of wild type.

  • PDF

Purification and Properties of Aspergillus ficuum Endoinulinase (Aspergillus ficuum 조효소액으로부터 Endoinulinase의 정제 및 특성)

  • Han, Sang-Bae;Ryu, Hyang-Suk;Rho, Min-Whan;Lee, Tae-Kyoo;Sohn, Hee-Suk;Woo, Soon-Ja;Uhm, Tai-Boong
    • Microbiology and Biotechnology Letters
    • /
    • v.19 no.2
    • /
    • pp.158-162
    • /
    • 1991
  • Endoinulinase was purified from a commercial inulin preparation produced by Aspergillus ficuum using ion exchange chromatography on CM-Sephadex C-50 and DEAE-Sepharose 6B, HPLC gel filtration on a Protein Pak 125 Colum and HPLC ion exchange chromatography on a TSK DEAE-5pw Column. The endoinulinase had a molecular weight of 72,000${\pm}$1,000 and was glycoprotein with 23 to 25% w/w sugar content. The enzyme was much more active on inulin with random cleavage mode than on sucrose and on palatinose: The ration of activity on inulin and sucrose (I/S ratio) was 10~14.

  • PDF

High-Level Expression of Aspergillus ficuum Acetyl Xylan Esterase Gene in Pichia pastoris, (Pichia pastoris에서 Aspergillus ficuum 유래 Acetyl Xylan Esterase 유전자의 과발현)

  • 임재명;김성구;박승문;남수완
    • Microbiology and Biotechnology Letters
    • /
    • v.30 no.4
    • /
    • pp.305-311
    • /
    • 2002
  • Acetyl xylan esterase gene (AXE) from Aspergillus ficuum was cloned and its Pichia expression plasmid, pPICZ$\alpha$C-AXE (4.6 kb), was constructed, in which the AXE gene was under the control of the AOXI promoter and connected downstream of mating factor u-1 signal sequence. The plasmid linearized by Sacl was integrated into the 5'AOXI region of the chromosomal DNA of P. pastoris. In the flask batch culture of P. pastoris transformant on methanol medium, the cell concentration and total AXEase activity reached at 6.0 g-dry cell weight/1 and 77 unit/ml after 36 h cultivation, respectively. In the fed-batch culture employing the optimized methanol and histidine feeding strategy, the cell concentration and total AXEase activity were significantly increased to about 97 g-dry cell weight/1 and 930 unit/ml. Most of AXEase activity (>90%) was found in the extracellular medium and the majority of extracellular protein (>80%) was AXEase enzyme (33.5 kDa). This result means that about 9.8 g/1 of AXEase protein was produced in the extracellular medium.

Studies on the Citric Acid Fermentation with Fungi (Part I) Isolation and Identification of Strains (사상균에 의한 구연산발효에 관한 연구 (제I보) 균주의 분리 및 동정)

  • 성낙계;김명찬;심기환;정덕화
    • Microbiology and Biotechnology Letters
    • /
    • v.8 no.1
    • /
    • pp.47-53
    • /
    • 1980
  • For the Purpose of studies on the citric acid fermentation, 579 strains of Aspergilli were isolated from natural sources of microorganisms. Out of them, the strains of M-80 and M-315 which produced relatively larger amount of citric acid than any others were selected after calling out an extensive screening test. The results obtained in light of the manual of Raper had been shown that the selected strains of M-80 and M-315 were identified as Aspergillus usamii mut. shirousamii, Aspergillus ficuum, respectively.

  • PDF

Comparative Enzymatic Hydrolysis of Bacillus amyloliquefaciens DS11 Phytase and Aspergillus ficuum Phytase in the Cannulated Pigs (Cannula를 장착한 돼지에서 Bacillus amyloliquefaciens DS11 Phytase와 Aspergillus ficuum Phytase의 역가 비교)

  • 장범수;박승춘;윤효인
    • Journal of Veterinary Clinics
    • /
    • v.18 no.1
    • /
    • pp.18-21
    • /
    • 2001
  • In this study, we assessed the efficacy of a novel B. amyloliquefacience DS11 phytase (DS11 phytase) and that of a commercial Aspergillus ficcum phytase (AF phytase) through their bioavailabilities of phytin-posphorus and -calcium in the diet using cannulated pigs. For the purpose of evaluating the efficacy of the phytases in pigs, we determined phosphorous concentrations from serum and feces, in addition to ingesta obtained from the cannula at the terminal ileum. As results, phosphorus concentration was lower in feces from DS11 group and BASF group by 17% and 10%, and higher in serum from the respective groups by 34% and 20%, as compared to the control group. Both phytases are evaluated to enhance phosphorus availability to the great extent. Calcium concentration of feces were lower in DS11 group and BASF group by 31% and 10%, than that in the control. Calcium concentration of serum was higher in DS11 phytase group by 4% but lower in AF phyase group by 3%, then that in the control group.

  • PDF

Expression of the Aspergillus niger var. awamori Phytase Gene in Pichia pastoris, and Comparison of Biological Properties

  • CHOI, JAE-MUN;DOO-SANG KIM;MOON-SICK YANG;HYUNG-RAK KIM;JAE-HO KIM
    • Journal of Microbiology and Biotechnology
    • /
    • v.11 no.6
    • /
    • pp.1066-1070
    • /
    • 2001
  • The PhyA gene, encoding myo-inositol hexakisphosphate phosphohydrolase in Aspergillus niger var. awamori (wild-type), was cloned and sequenced. The cDNA was overexpressed by a multicopy gene expression system in Pichia pastoris KM71. Recombinant, wild-type and commercial phytase from Aspergilus ficuum NRRL 3135 (Natuphos) were purified. The PhyA gene of Aspergillus niger var awamori showed perfect homology to the phytase of Aspergillus ficcum and $97\%$ homology to A. niger var awamori (L02421). Wild-type phytase was highly glycosylated and more thermostable than the other two, while deglycosylated farms of three phytases showed identical molecular weight, 507 kDa. After heating at $80^{\circ}C$, wild-type, commercial, and recombinant phytases retained $57\%, 32%,\;and\;8\%$ of their original activities, respectively. In conclusion, glycosylation plays a key role in the thermostability of phytase and its enzymatic characterization.

  • PDF

Cloning and Heterologous Expression of Acetyl Xylan Esterase from Aspergillus ficuum

  • Jeong, Hye-Jong;Park, Seung-Mun;Yang, Mun-Sik;Kim, Dae-Hyeok
    • 한국생물공학회:학술대회논문집
    • /
    • /
    • pp.153-156
    • /
    • 2000
  • Xylan, the major hemicellulose component of many plants, occurs naturally in a partially acetylated form and lignin, the most resistant component in plant cell wall degradation, is also attached to ${\beta}-1,4-linked-D-xylose$ backbone through the ester linkage. Esterases are required to release the esterified substituent and acetyl esterases are important in the complete degradation of acetylated polysaccharides, like pectins and xylans. The gene(Axe) encoding acetyl xylan estarase(AXE) was isolated from genomic ${\lambda}$ library from Aspergillus ficuum. Nucleotide sequencing of the Axe gene indicated that the gene was separated with two intervening sequences and the amino acid sequence comparison revealed that it was closely related to that from A. awamori with the 92 % indentity. Heterologous expression of AXE was conducted by using YEp352 and Saccharomyces cerevisae 2805 as a vector and host expression system, respectively. The Axe gene was placed between GAL1 promoter and GAL7 terminator and then this recombinant vector was used to transform S. cerevisiae 2805 strain. Culture filtrate of the transformed yeast was assayed for the presence of AXE activity by spectrophotometry and, comparing with the host strain, four to five times of enzyme activity was detected in culture filtrate of transformed yeast.

  • PDF