• Title/Summary/Keyword: enzyme-resistant starch

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Preparation of Indigestible Dextrin from Pyrodextrin (열처리 덱스트린을 이용한 난소화성 덱스트린의 제조)

  • Woo, Dong-Ho;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.618-628
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    • 2000
  • The indigestible dextrin I was prepared by hydrolyzing pyrodextrin with thermostable ${\alpha}-amylase$. The mean values of indigestible fraction and dieatry fiber of indigestible dextrin I prepared from yellow dextrin were 50.0% and 25.0%, respectively. Also the indigestible dextrin II was prepared by removing low molecular weight saccharides containing glucose with ethanol from enzyme hydrolysate of pyrodextrins. Over 80% of glucose and maltose in initial enzyme hydrolysate were removed, therefore the indigestible fraction and dietary fiber of the indigestible dextrins increased. The indigestible dextrin from ethanol precipitate of enzyme hydrolysate of yellow dextrin by ${\alpha}-amylase$ and amyloglucosidase showed a higher contents of indigestible fraction and dietary fiber than ethanol precipitates by any other enzyme combination, and its mean values were 83.6% and 62.8%, respectively. Consequently, it was found that the indigestible dextrins which are resistant to starch-hydrolysing enzyme can be easily prepared from pyrodextrin, and presumed that they can perform physiological functions as soluble dietary fiber.

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Purification and Characterization of Alkali-resistant Amylases from Pseudomonas sp. (Pseudomonas sp.로부터 알칼리내성 amylase의 정제 및 특성 확인)

  • Lee, Jeong-Eun;Jhon, Deok-Young
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.70-75
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    • 2008
  • Two extracellular amylase isozymes were purified and characterized from alkalophilic Pseudomonas sp. KFCC 10818 for the production of maltooligosaccharides. The molecular weights of the homogeneous proteins were 50 kDa and 75 kDa, respectively. The 50 and 75 kDa amylases showed optimum temperatures at 35 and $40^{\circ}C$, respectively. The optimum pH of the enzymes ranged from pH 6-8, and the enzymes were resistant to an alkaline condition of pH 12. Via the enzyme's actions, the final products from maltooligosaccharides or soluble starch were maltose and maltotriose. Calcium was a potent activator of the 50 kDa amylase. Finally, the N-terminal amino acid sequences of the 50 and 75 kDa amylases were QTVPKTTFV and DTVPGNAFQ, respectively.

Purification and Characterization of Cyclodextrin Glucanotransferase from Paenibacillus sp. JK-12

  • Kang, Yong;Kim, Sung-Koo;Jun, Hong-Ki
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.310-316
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    • 2002
  • Extracellular cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JK-12 was purified through sev-eral purification steps consisting of ammonium sulfate precipitation and chromatographies on DEAE-sephadex A-50 and Mono QIM HR5/5. The purified CGTase exhibited a single band on SDS-PAGE and was estimated to be approximately 82 kDa. The isoelectric point of the enzyme was 7.2 as determined by isoelectric focusing. The CGTase from Paenibacillus sp. JK-12 had a transglucosylation activity at the C-2 position of L-ascorbic acid. The optimum pH and temperature for the CGTase activity were 8.0 and 5$0^{\circ}C$, respectively. The enzyme activity was stable from pH 6.0 to 9.() and at temperatures up to 55$^{\circ}C$ at pB 8.0, having 80% residual activity. The activity of the CGTase was strongly resistant to metals such as A $g^{+}$ and $Ba^{2+}$ but slightly inhibited by H $g^{+}$, N $i^{2+}$ and $Mg^{2+}$. The enzymeproduced $\alpha$ -cyclodextrin ($\alpha$-CD) and $\beta$-CD as the main products from starch, but not ${\gamma}$-CD.X>-CD.

Purification and Properties of Glucoamylase form Yeast Candida tsukuaensis

  • Kim, Sanga-Moon;Bai, Suk;Chung, Hee-Young;Park, Jong-Chun;Lee, Jin-Jong;Kim, Dong-Ho;Song, Myoung-Hee;Chun, Soon-Bai
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.519-523
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    • 1992
  • The glucoamylase of Candida tsuubaensis was purified to homogeneity form culture filtrate by means of ultrafiltration, Sephacryl S-200 gel filtration and Sp-Sephadex C-50 chromatography. The purified enzyme was a glycoprotein with a molecular mass of approximately 50 kDa, which was a monomeric protein. Km values were 5.8 mg/ml for soluble starch and 0.04 mM for maltose. Glucoamylase also released only glucose from both pullulan and isomaltose. The analysis of amino acid composition revealed that the enzyme contained a high content of acidic and polar amino acids. In addition, Western blotting analysis indicates that C. tsukubaensis glucoamylase is resistant to glucose repression.

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Development of a Mixed-culture Fermentation Process and Characterization for New Oligosaccharides and Dextran Using Lipomyces starkeyi and Leuconostoc mesenteroides (Lipomyces starkeyi와 Leuconostoc mesenteroides의 혼합배양에 의한 올리고당과 Dextran의 생성 및 생성당의 특성 연구)

  • 허수진;김도만;이인수;장판식
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.304-310
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    • 1999
  • We have developed a new process for the production of new structure oligosaccharides using the mixed-culture fementation of Lipomyces starkeyi KSM22 and leuconostoc mesenteroides B-512FMCM.L.starkeyi KSM22 produces a novel DXAMase(an enzyme containing both dextranase and amylase activities). It hydrolyzes the soluble starch and dextran. The hydrolyzates were used as acceptors for dextransucrase of L.mesenteroides to synthesize the new oligosaccharides(NOS). In fermentation, as the concentration of sucrose was increased from 9%(w/v) to 15%(w/v), the yields of dextran(sum of dextran I, MW=66kD, and dextran II, MW=21kD) was increased from 12.7% to 42.5%, and NOS was increased from 3.9% to 5.2% of the theoretical, respectively. The NOS of dp(degree of polymerization) 5 and over was increased from 33.1% to 58.3% of the total NOS. The NOS showed heat resistant up to 12$0^{\circ}C$ and was stable at pHs ranged from 2 to 6. The NOS decreased the pH changes in the culture of S. mutans, and also showed inhibitory effects on the growth of S. aureus or S. typhimurium.

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Qualitative and quantitative PCR detection of insect-resistant genetically modified rice Agb0101 developed in korea (해충저항성 유전자변형 벼 Agb0101에 대한 PCR 검정)

  • Shin, Kong-Sik;Lee, Jin-Hyoung;Lim, Myung-Ho;Woo, Hee-Jong;Qin, Yang;Suh, Seok-Cheol;Kweon, Soon-Jong;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.18-26
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    • 2013
  • Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.