• Title/Summary/Keyword: Amylolytic Activity

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Diurnal Fluctuations of Saprophytic Bacterial distribution and Their Extracellular Enzyme Activities in the Overlying Waters of Sediment of the Yellow Sea near Daesan, Korea (대산인근 해역에서 간만조에 따른 종속영양세균의 일일 분포와 세포외 효소 활성력의 변화)

  • Lee, Geon-Hyoung;Gang-Guk Choi;Chun-Bong Baek
    • The Korean Journal of Ecology
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    • v.18 no.3
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    • pp.409-418
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    • 1995
  • As a part of studying the function and structure of the mudflat environment of the Yellow Sea, seawater samples in the overlying waters of sediment near Daesan were collected every hour on March 29 (spring tides) and on April 5 (neap tides), 1995 to study the diurnal distribution of aerobic saprophytic bacteria and their extracellular enzyme activities. The diurnal distribution of aerobic saprophytic bacteria ranged from 1.0 X $10^{2}$ to 7.07 X $10^{3}$ cfu /ml at spring tides and from 1.0 X $10^{2}$ to 8.3 X $10^{3}$ cfu /ml at neap tides. The diurnal variations of aerobic saprophytes at the suface waters were greater than those of middle and bottom waters. However, th diurnal fluctuation of saprophyte numbers at spring tides showed no significant difference compared with that at neap tides. The numbers of three physiological groups of aerobic hacteria (proteolytic, lipolytic and amylolytic bacteria) at the surface waters during spring and neap tides were lower than those at the middles and bottom waters. The diurnal variations of five extracellular enzyme activities at the surface waters during the survey period showed lower values than those at the middle and botton waters. Among the measured extracellular enzyme activities, phosphatase showed the highest. However, the activities of amylase, chitinase and cellulase showed a similar tendency.

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$\alpha$-Amylase와 Glucoamylase를 동시에 분비하는 배수체 재조합효모에 의한 전분기질로 부터의 에탄올 생산

  • Park, Sun-Young;Kim, Min-Soo;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.604-612
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    • 1996
  • To improve the fermentation characteristics of the haploid starch-fermenting recombinant yeast strain K114/YIpMS$\Delta$R(LEU2/URA3) secreting both $\alpha$-amylase and glucoamylase was rare-mated with polyploid industrial yeast Saccharomyces sp. K35. The K35 strain had good fermentation-characteristics such as ethanol-tolerance, high temperature and sugar-tolerance, and high fermentation rate. Among the resulting 66 hybrids, the best strain RH51 was selected. The RH51 exhibited amylolytic activity of K114/YIpMS$\Delta$R(LEU2/URA3) as well as ethanol and sugar tolerance of K35. The optimum temperature of hybrid RH51 for starch fermentation was 34$\circ$C which was same as that of K35 but different from that (30$\circ$C) of K114/YIpMS$\Delta$R(LEU2/URA3). The optimum pH was 5.0. The optimum size of inoculum was 2% as the pellet (w/v) of yeast cells. The hybrid strain RH51 produced 7.0% ethanol (w/v) from 20% (w/v) soluble starch while K35 formed almost no ethanol, 0.3% (w/v). RH51 strain produced 7.5% (w/v) ethanol after 8 days in a 2.5 l fermenter containing 800 ml of 20% (w/v) soluble starch. The residual starch content in the fermentation medium was 1.68% (w/v), and therefore almost all the starch was fermented completely.

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Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Isolation and Characteristics of Bacteriocin-producing Bacteria from the Intestine of Duck for Probiotics (오리로부터 박테리오신을 생산하는 프로바이오틱 미생물의 분리 및 특성)

  • Shin, M.S.;Han, S.K.;Ji, A.R.;Ham, M.R.;Kim, K.S.;Lee, W.K.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.621-632
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    • 2007
  • The aim of this study was to isolate and characterize bacteriocin-producing bacteria from the intestine of duck to use as probiotics for livestock. A total of 416 strains were isolated from the small intestine and cecum of ducks and 13 isolates were finally selected after determinging inhibitory activity against pathogenic indicators by spot-on-lawn method. The selected strains were identified as Lactobacillus salivarius JWS 58, Lactobacillus plantarum JWS 1354, Pediococcus pentosaceus JWS 939, 7 strains of enterococci, and 3 strains of Escherichia coli. Lact. salivarius JWS 58, Ent. faecium JWS 833, and Ped. pentosaceus JWS 939 showed a strong inhibitory activity against Listeria monocytogenes. E. coli JWS 108 inhibited the growth of E. coli and Staphylococcus aureus. Lact. salivarius JWS 58 strain survived almost 50% in pH 2.5 phosphate buffer for 2 hr. Ped. pentosaceus JWS 939 and Lact. plantarum JWS 1354 showed strong amylolytic activity. These results suggest that a combination of bacteriocins or multispecies probiotics of the selected strains has a strong potential of alternative to antibiotics in livestock production.

Cloning of Isoamylase Gene of Pectobacterium carotovorum subsp. carotovorum LY34 and Identification of Essential Residues of Enzyme (Pectobacterium carotovorum subsp. carotovorum LY34에서 Lsoamylase 유전자 클로닝 및 효소 활성의 필수 잔기 확인)

  • Cho, Kye-Man;Kim, Eun-Ju;Math, Renukaradhya K.;Asraful Islam, Shah Md.;Hong, Sun-Joo;Kim, Jong-Ok;Shin, Ki-Jae;Lee, Young-Han;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1182-1190
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    • 2007
  • The gene encoding for isoamylase of the Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 was cloned and expressed into Escherichia coli $DH5{\alpha}$. Isoamylase catalyzes the hydrolysis of ${\alpha}-1,6-glycosidic$ linkages specifically in amylopectin, glycogen, and derived oligosaccharides, while the enzyme did not hydrolyze ${\alpha}-1,4-glycosidic$ linkages of amylose. The isoamylase gene (glgX) had an open reading frame of 1,977 bp encoding 658 amino acid residues with a calculated molecular weight of 74,188 Da. The molecular weight of the enzyme was also estimated to be 74 kDa by activity staining of a SDS-PA gel. The mature GlgX had a calculated pI of 4.91. Isoamylase from Pcc LY34 had 70% amino acid identity with isoamylase from Pectobacterium chrysanthemi and contained the four regions conserved among all amylolytic enzymes. The isoamylase was optimally active at pH 7.0 and $40^{\circ}C$. GlgX was $Ca^{2+}-dependent$. The changes of Asp-335, Glu-370, and Asp-442 into Ala, respectively, using site-directed mutagenesis techniques showed that three residues are essential to isolamyalse (GlgX) activity. The sequences around those residues were highly conserved in isoamylase of different origins and GlgX of the glg operon in glycongen biosynthesis.

Fermentation and Quality Characteristics of Cheonggukjang with Addition of Dropwort (Oenanthe javanica D.C.) Powder (미나리 분말을 첨가한 청국장의 발효 및 품질특성)

  • Lee, Shin-Ho;Kim, Jin-Hak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1133-1138
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    • 2013
  • Characteristics of Cheonggukjang with addition of different dropwort (Oenanthe javanica D.C.) powders were investigated. The selected strain, with proteolytic, amylolytic, and antimicrobial activity, was identified as B. subtilis RS-9, using 16S rRNA analysis. The Cheonggukjang was prepared with cooked soybean without dropwort (Control), 0.5% raw dropwort powder (DW0.5), and 1% raw dropwort powder (DW1), 0.5% steamed dropwort powder (SDW0.5), and 1% steamed dropwort powder (SDW1) were added, respectively. The changes in pH of Cheonggukjangs with addition of dropwort powder were lower than those of control during fermentation for 72 hr at $40^{\circ}C$. The total aerobes of the various Cheonggukjangs reached 8.88 (control), 8.82 (DW0.5), 8.70 (DW1), 8.85 (SDW0.5), and 8.75 (SDW1) log CFU/mL after fermentation for 72 hr at $40^{\circ}C$, respectively. The amino nitrogen and viscous substance contents of different dropwort powders added to Cheonggukangs were lower than those of control. The total polyphenol contents and ABTS radical scavenging ability of various Cheonggukjangs were increased by addition of dropwort powder and fermentation. The polyphenol contents and ABTS radical scavenging ability of SDW1 were $590.24{\mu}g/mL$ and 82.16% and showed the highest value among tested Cheonggukangs. The sensory quality of DW0.5 was higher in taste and overall acceptability, compared with other groups.

Quality characteristics and preparing of solid starter using fungal strains for Takju (탁주용 곰팡이 고체종국 제조 및 품질 특성)

  • Baek, Chang-Ho;Baek, Seong Yeol;Mun, Ji-Young;Choi, Han-Seok;Kang, Ji-Eun;Jung, Seok-Tae;Yeo, Soo-Hwan
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.797-803
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    • 2016
  • In this study, we investigated the effect of fermentation conditions on the amylolytic and proteolytic activities of Aspergillus luchuensis strain 74-5 and Aspergillus oryzae strain 75-2, which are used in the preparation of the starter culture, for Takju (Korean traditional rice wine). The starter culture was optimized using different conditions, such as inoculum size, inoculation temperature, and incubation time. The enzyme activities under each condition were measured. In the A. luchuensis strain 74-5 starter culture, the ${\alpha}-amylase$ and glucoamylase activities increased, however the activity of acidic protease decreased as the diluent to starter culture ratio increased. In the A. oryzae 75-2 starter culture, all enzyme activities were maintained at a higher level even at 5% inoculation ratio. Higher enzyme activities were observed in the middle range of inoculation temperature (35, $40^{\circ}C$), than in the lower range (20, $30^{\circ}C$). Enzyme activity in the starter culture varied with incubation time, however it was the highest at 144 and 120 hr, respectively, for A. luchuensis strain 74-5 and A. oryzae strain 75-2. The spore count of the starter culture was approximately $2{\times}10^7$ during fermentation, out of which contamination by aerobic bacteria was about $3{\times}10^3$. The results suggested that the starter culture of each strain could be used as an inoculum for fermentation. However, we needs to conduct further research for the selection of suitable diluting agents as well as drying methods to reduce the contamination by aerobic bacteria, while retaining the enzyme activity.

Mitotic Stability of Heterologous $\alpha$-Amylase Gene in Starch-Fermenting Yeast (전분발효 효모에서의 외래 $\alpha$-Amylase 유전자의 세포분열시 안정성 증진)

  • Kim, Jung-Hee;Kim, Keun;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.271-279
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    • 1994
  • To develop a yeast strain which stably secretes both $\alpha$-amylase and glucoamylase and therefore is able to convert starch directly to ethanol, a mouse salivary $\alpha$-amylase cDNA gene with a yeast alcohol dehydrogenase I promoter has been introduced into the cell of a Saccharomyces diactaticus hybrid strain secreting only glucoamylase. To secrete both enzymes more stably without loss of the $\alpha$-amylase gene during a cell-multiplication, an integrating plasmid vector containing $\alpha$-amylase gene was constructed and introduced into the yeast cell. The results showed that the linearized form of the integrating vector was superior in the transformation efficiency and the rate of the expression of the $\alpha$-amylase gene than the circular type of the vector. The yeast transformant having a linearized plasmid vector exhibited higher mitotic stability than the yeast transformant habouring episomat plasmid vector. The transformant containing the linearized vector producing both $\alpha$-amylase and glucoamylase exhibited 2-3 times more amylolytic activity than the original untransformed strain secreting only glucoamylase.

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Fungal Microflora on Korean Home-made Meju (재래식(在來式) 메주에 분포(分布)하고 있는 진균(眞菌)에 관한 조사(調査) 연구(硏究))

  • Park, Kyoung-Ja;Kim, Young-Mi;Lee, Bae-Ham;Lee, Bok-Kwon
    • The Korean Journal of Mycology
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    • v.5 no.1
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    • pp.7-12
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    • 1977
  • For this study 3 samples of the home-made meju and 3 samples of the improved meju were selected according to their characteristics. A total of 25 strains of true fungi were isolated from the samples of the home-made meju and identified by the Key of Alexopoulos and Raper, using a strain Aspergillus oryzae A-113 as a control. Amylolytic and proteolytic enzyme activities of the isolated strains were investigated ana the results obtained were as follows. 1. The 25 strains from the home-made meju were identified into 2 Aspergillus oryzae, 14 Asp. flavus, 6 Penicillum spp. 1 Candida sp 1 Spicaria sp and 1 Rhizopus sp. 2. The 3 strains from the improved meju were all identified as Aspergillus oryzae. 3. Aspergillus flavus, A-B, from the home-made meju was found to he the strongest strain in ${\alpha}-amylase$ activity and also to be similar to the strains of Aspergillus orzae from improved meju. 4. Aspergillus flavus, A-7, from the home-made meju was found to be the strogest strain in ${\beta}-amylase$ activity and stronger than that from the improved meju. 5. Aspergillus flavus B-3, was found to be the strongest strain in protease activity and stronger than that from the improved meju. 6. Some of the strains from the home-made meju turned out to be harmful strains, such as Penicillium spp. which secrete antibiotics, Asp. flavus which secretes mycotoxin, Candida sp which causes skin diseases, Spicaria sp. which is a insect pathogen. 7. Rhizopus sp was also found but it has not been proved to be harmful.

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