• Title/Summary/Keyword: ${\beta}$-glucanase

Search Result 187, Processing Time 0.024 seconds

Response of ${\beta}-Glucanases{\;}to{\;}GA_3$ in Barley Aleurone Layers (보리 호분층에서 $(1-3)-{\beta}-glucanase{\;}$${\;}(1-3,1-4)-{\beta}-glucanase$${\;}GA_3$에 대한 반응)

  • Song Joong, Yun;Ill Min, Chung
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.40 no.2
    • /
    • pp.250-254
    • /
    • 1995
  • Isolated barley aleurone layers were used to examine response of (1-3)- and $(1-3,1-4)-{\beta}-glucanases{\;}to{\;}GA_3$. Protein content and levels of (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ increased in the presense of added $GA_3$. However, (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ showed different response to $GA_3$ in their production and secretion patterns. $(1-3,1-4)-{\beta}-glucanases$ showed higher increase in enzyme activity than $(1-3)-{\beta}-glucanase$ in the early stage of$GA_3$treatment. Secretion of enzyme by $GA_3$ into the surrounding medium was more enhanced in $(1-3,1-4)-{\beta}-glucanases$ than in $(1-3)-{\beta}-glucanase$, The differential response of the enzymes might be related to the physiological role of the enzymes in germination of barley grain.

  • PDF

Synthesis and Secretion of the Endo-$\beta$-l,4-Glucanase from Bacillus subtilis in Industrial Yeast Strain (산업용 효모에서 Bacillus subtilis Endo-$\beta$-1,4-Glucanase의 생합성 및 분비)

  • 박용준;이영호;백운화;강현삼
    • Microbiology and Biotechnology Letters
    • /
    • v.19 no.4
    • /
    • pp.348-355
    • /
    • 1991
  • DNA segment encoding $\beta$-1, 4-glucanase of Bacillus subtilis was fused in frame to mouse $\alpha$-amylase signal sequence behind the alcohol dehydrogenase isoenzyme I gene (ADHI) promoter of the yeast expression vector pMS12. To enhance the expression level of the $\beta$glucanase gene in yeast, transcription terminator sequence iso-1-cytochrome c gene (CYCI) was inserted into the recombinant plasmid. The transformants harbouring such recombinant plasmids secreted $\beta$-glucanase into the culture medium. The expresstion level of the $\beta$-glucanase gene was increased about 2-fold caused by inserting the terminator. The amount of the secreted $\beta$-glucanase in culture medium was approximately 60% of the total quantity synthesized.

  • PDF

Direct Detection of (1-3)-$\beta$-Glucanase Isozymes in Isoelectrofocusing Gels Using a Dye -Labeled Substrate (염료착색 기질을 이용한 IEF gel에서(1-3)-$\beta$-glucanase 동위효소의 검출)

  • Yun, Song-Joong;Lee, Myong-Chul;Kwon, In-Sook;Kim, Tae-San;Go, Seung-Joo
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.39 no.2
    • /
    • pp.121-127
    • /
    • 1994
  • A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

  • PDF

Enhancement of β-1,3-Glucanase Activity by Sequential δ-Sequence Mediated Integration in Saccharomyces cerevisiae (출아효모에서 연속적 δ-sequence 삽입유도에 의한 β-1,3-glucanase 활성 증가)

  • Kim, Min-Jung;Kim, Yeon-Hee
    • Journal of Life Science
    • /
    • v.24 no.10
    • /
    • pp.1046-1054
    • /
    • 2014
  • Beta-1,3-glucanase is widely used in various biotechnological and industrial processes, with over-production required to enable versatile utilization. We examined the overexpression of ${\beta}$-1,3-glucanase (EXGA) from Aspergillus oryzae using ${\delta}$-sequence-mediated integration. We constructed $pRS{\delta}$-exgA and $pRS{\delta}K$-exgA plasmids for integration of the EXGA gene into various chromosomes of Saccharomyces cerevisiae. These plasmids contain the ADH1 promoter for constitutive expression, a signal sequence (exoinulinase signal sequence [INU1 s.s]) for secretory production, and a ${\delta}$-sequence for integration of ${\beta}$-1,3-glucanase. The $pRS{\delta}$-exgA plasmid was transformed into the S. cerevisiae $BY4742{\Delta}exg1$ strain, and ${\beta}$-1.3-glucanase was stably overexpressed and secreted. Another plasmid, $pRS{\delta}K$-exgA, was introduced into the S. cerevisiae $BY4742{\Delta}exg1$ (YKY082) strain, and overexpression of ${\beta}$-1,3-glucanase was examined by inducible integration under geneticin selection. The activity of ${\beta}$-1,3-glucanase increased in accordance with a rise in the geneticin concentration, with 0.8 mg/ml of geneticin suitable for overexpression of ${\beta}$-1,3-glucanase. Subsequently, $pRS{\delta}K$-exgA was repeatedly transformed for sequential ${\delta}$-integration. The activity of ${\beta}$-1,3-glucanase reached about 0.063 unit/ml/$OD_{600}$, 0.095 unit/ml/$OD_{600}$, 0.131 unit/ml/$OD_{600}$ and 0.165 unit/ml/$OD_{600}$ by the first, second, third, and fourth round of integration, respectively. According to the increase in the activity of ${\beta}$-1,3-glucanase by sequential ${\delta}$-integration, the copy number (integration rate) of the EXGA gene also increased in various chromosomes. These results suggest that recombinant ${\beta}$-1,3-glucanase activity can be sequentially increased by repeated ${\delta}$-sequence integration.

Characteristics of $endo-{\beta}-1,3-glucanase$ from green malt (녹맥아에서 추출한 $endo-{\beta}-1,3-glucanase$의 효소학적 성질)

  • Son, Bong-Soo;Sung, Nack-Kie
    • Applied Biological Chemistry
    • /
    • v.35 no.3
    • /
    • pp.165-169
    • /
    • 1992
  • Two types of $endo-{\beta}-1,3-glucanases$ were purified from green malt and their basic characteristics were studied. Molecular weights of glucanase I and glucanase II were estimated, by electrophoresis, to be 35,000 and 28,000, respectively. Purified glucanase I and II showed the highest activity at pH $5.0{\sim}7.0$ and $5.0{\sim}8.0$, respectively. The optimal temperature of purified glucanase I and II was $40^{\circ}C$. Purified glucanase I and glucanase II were stable at $40^{\circ}$ for 60 min and at $50^{\circ}$ for 30 min. All enzymes were inactivited by $AgNO_3$ and $HgCl_2$ while those were not activated by various compounds tried. Km values of glucanase I and II were 1.03 mg/ml, 1.20 mg/ml, respectively.

  • PDF

(1-3, 1-4)-$\beta$-Glucan and Starch Contents and Their Hydrolytic Enzyme Activities in Developing Barley Kernels (등숙 중인 보리 종실중 (1-3, 1-4)-$\beta$-Glucan과 전분 함량 및 이들의 가수분해효소 활성)

  • 윤성중;박상래;유남희
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.42 no.4
    • /
    • pp.403-409
    • /
    • 1997
  • To obtain information on the accumulation of (1-3, 1-4)-$\beta$-glucans during kernel maturation, (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities were determined in developing kernels of the two Korean cooking barley varieties, Neulssalbori and Saessalbori. (1-3, 1-4)-$\beta$-Glucan contents in kernels at 5 and 10 days after anthesis(DAA) were very low and the contents increased rapidly in kernels at 15 to 25 DAA. (1-3, 1-4)-$\beta$-Glucan content in kernels at harvest was about 3.5 to 4% of kernel dry matter. (1-3, 1-4)-$\beta$-Glucanase activities were relatively higher in younger kernels but the levels of the activity were very low compared with those in germinating kernels. A significant negative correlation was observed between (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities. Low levels of (1-3, 1-4)-$\beta$-glucanase activites in kernels at 15 to 30 DAA, however, may indicate that (1-3, 1-4)-$\beta$-glucanases have little effect on the final content of (1-3, 1-4)-$\beta$-glucans in barley kernels. Starch contents and $\alpha$-amylase activities were also determined in developing barley kernels. Starch contents increased rapidly as kernels matured and the content at harvest was about 60% of kernel dry matter. Relativley higher levels of $\alpha$-amylase activities in kernels at the earlier developmental stage decreased rapidly as kernels matured.

  • PDF

Isolation and Properties of a Protein, RCG-2, Having Chitinase, ${\beta}-1,3-Glucanase$ and Lysozyme Activities from Rice Leaves (Chitinase, ${\beta}-1,3-glucanase$ 및 lysozyme 효소활성을 보유한 벼잎 산성단백질 RCG-2)

  • Um, Sung-Yon;Kim, Su-Il
    • Applied Biological Chemistry
    • /
    • v.37 no.1
    • /
    • pp.49-55
    • /
    • 1994
  • An acidic protein, RCG-2, containing chitinase and ${\beta}-1,3-glucanase$ activity conccurrently was purified from rice leaves by chromatofocusing and gel slicing. The purified enzyme gave a single band on polyacrylamide gel electrophoresis and its molecular weight was appeared to be 29.7 kd using SDS-PAGE. This enzyme also had lysozyme activity. The optimal temperature for both enzyme activities was $40^{\circ}C$, optimal pH were 4.0 for chitinase activity and 7.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 7.86 mM and $0.025\;{\mu}M/min.$, and those for ${\beta}-1,3-glucanase$ were 5.95 mM and $0.16\;{\mu}M/min.$ respectively. TLC analysis of the enzyme hydrolysates of chitooligosaccharides indicated that this enzyme acts as endochitinase.

  • PDF