• Title/Summary/Keyword: hydrolysis yield

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Enzymatic Biotransformation of Ginsenoside Rb2 into Rd by Recombinant α-L-Arabinopyranosidase from Blastococcus saxobsidens

  • Kim, Ju-Hyeon;Oh, Jung-Mi;Chun, Sungkun;Park, Hye Yoon;Im, Wan Taek
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.391-397
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    • 2020
  • In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenoside-converting Blastococcus saxobsidens that was cloned and expressed in Escherichia coli BL21 (DE3), and then applied it in the biotransformation of ginsenoside Rb2 into Rd. The gene, termed AbpBs, consisting of 2,406 nucleotides (801 amino acid residues), and with a predicted translated protein molecular mass of 86.4 kDa, was cloned into a pGEX4T-1 vector. A BLAST search using the AbpBs amino acid sequence revealed significant homology with a family 2 glycoside hydrolase (GH2). The over-expressed recombinant AbpBs in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinopyranose moiety attached to the C-20 position of ginsenoside Rb2 under optimal conditions (pH 7.0 and 40℃). Kinetic parameters for α-L-arabinopyranosidase showed apparent Km and Vmax values of 0.078 ± 0.0002 μM and 1.4 ± 0.1 μmol/min/mg of protein against p-nitrophenyl-α-L-arabinopyranoside. Using a purified AbpBs (1 ㎍/ml), 0.1% of ginsenoside Rb2 was completely converted to ginsenoside Rd within 1 h. The recombinant AbpBs could be useful for high-yield, rapid, and low-cost preparation of ginsenoside Rd from Rb2.

A Mass Preparation Method of (+)-Decursinol from the Roots of Angelica gigas (참당귀(Angelica gigas) 뿌리 추출물로부터 (+)-Decursinol의 대량 제조 방법)

  • Lee, Jee-Hyun;Choi, Yong-Seok;Kim, Joo-Hwan;Jeong, Hye-Gwang;Kim, Dong-Hee;Yun, Mi-Yong;Kim, Jin-Sook;Lee, Sang-Han;Cho, Soo-Hyun;Shen, Gui-Nan;Kim, Eui-Geom;Jin, Wen-Yi;Song, Gyu-Yong
    • YAKHAK HOEJI
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    • v.50 no.3
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    • pp.172-176
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    • 2006
  • The purpose of this study is to develop a mass preparation method of (+)-decursinol from Angelica gigas roots. Recently; it has been shown that (+)-decursin, (+)-decursinol angelate and its analogues, isolated from Angelica gigas roots, exhibit various biological activities such as antitumor, antibacterial and neuroprotective activities. The contents of these compounds, ester form of (+)-decursinol, is very high in the Angelica gigas roots, whereas the content of (+)-decursinol itself is very low Therefore, (+)-decursinol which can be used as starting material to synthesize various its analogues was easily prepared from decursin analogues in the Angelica gigas roots. In order to achieve such aim, the Ether-Fr. of the ethanol exact of Angelica gigas roots was hydrolysed with various alkalis and solvents. As a result, the order of (+)-decursinol preparation was 1) NaOH, KOH, 2) $K_2CO_3$, and 3) $NaHCO_3$ as alkali. Also, the yield of (+)-decursinol was higher in diethyl ether than any other solvent conditions. From 1 kg of dried Angelica gigas roots, we could obtain 27.4 g of (+)-decursinol as a pure white solid.

Effects of Artificial Stomach Fluid and Digestive Enzymes on the Aglycone Isoflavone Contents of Soybean and Black Bean (Rhynchosia Molubilis : Yak-Kong) (대두와 쥐눈이콩의 비배당체 이소플라본 함량에 대한 인공위액과 소화효소 처리효과)

  • 강순아;장기효;조윤희;홍경희;서지혜;조여원
    • Journal of Nutrition and Health
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    • v.36 no.1
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    • pp.32-39
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    • 2003
  • Phytoestrogens, especially soy-derived isoflavones, are receiving great scrutiny as a food supplement for preventing hormone dependent disease such as postmenopausal osteoporosis. Their beneficial effects are derived from aglycone form of isoflavones, such as daidzein, genistein or glycitein. In contrast to the common usage of soybean, black bean (Rhynchosia Molubilis : Yak-kong) has been used as a supplement for preventing postmenopausal osteoporosis in oriental medicine. To investigate the effects of the saliva, artificial stomach fluid, and digestive enzymes on the conversion of glycosidic isoflavone to aglycone form, soybean and black bean were extracted with 70% methanol and freeze-dried. The recovery yield of methanol extracts of black bean was 14.1% which was higher than that of soybean, 13.5%. In terms of total isoflavones, we routinely obtained larger amount of isoflavones from black bean than those from soybean. By incubating methanol extracts of soybean and black bean with IN HCI for 180 min, the proportions of aglycones relative to the total isoflavone were significantly increased (32.4% and 52.4%, respectively). In vitro conversion, digestive enzymes ($\beta$-glucosidase and $\alpha$-glucosidase) may hydrolyze glycosidic bond of isoflavone more effectively than saliva or artificial stomach fluid did. It seems to say that the activity of $\beta$-glucosidase was higher than those of $\alpha$-glucosidase. The rate of conversion of glucoside form to aglycone form in black bean and soybean was low in physiological condition (pH) tested, although the enzymatic hydrolysis of glucoside was active. These results demonstrated that the composition of aglycone in food may be the important factors in terms of the bioavailability of isoflavones. (Korean J Nutrition 36(1): 32-39, 2003)

Microalgae Removal and Energy Production by Combined Electro-flotation and Anaerobic Hydrogen Fermentation Processes (전기부상과 혐기성 수소 발효 공정의 결합을 통한 미세조류 제거 및 에너지 생산)

  • Lee, Chae-Young;Na, Dong-Chae;Choi, Jae-Min;Kang, Doo-Sun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.20 no.3
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    • pp.83-88
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    • 2012
  • The algal bloom, resulting from eutrophication, has caused serious water quality problems in river and lake. Therefore, it has to be removed by any means including physicochemical or biological treatment for preserving water quality. This study was conducted to investigate the microalgae removal and energy production using combined electro-flotation and anaerobic hydrogen fermentation processes. The result showed that algae removal efficiency based on chlorophyll a removal increased with the current. At a current of 0.6A, the maximum microalgae removal efficiency of 95.9% was achieved. The treatability of anaerobic hydrogen fermentation was investigated to recover energy from microalgae removed by electro-flotation. The ultimate hydrogen yields of algae before and after ultrasonic pretreatment were 17.3 and 61.1 ml $H_2/g$ dcw(dry cell weight), respectively. The ultrasonic pretreatment of algae led to 3.4-fold higher $H_2$ production due to the increase of hydrolysis rate.

Microbial Community Dynamics in Batch High-Solid Anaerobic Digestion of Food Waste Under Mesophilic Conditions

  • Yi, Jing;Dong, Bin;Xue, Yonggang;Li, Ning;Gao, Peng;Zhao, Yuxin;Dai, Lingling;Dai, Xiaohu
    • Journal of Microbiology and Biotechnology
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    • v.24 no.2
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    • pp.270-279
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    • 2014
  • Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g $VS_{removal}$. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

Enzymatic Modification of Sardine Protein Concentrate (정어리 분말(粉末) 단백질(蛋白質)의 효소적(酵素的) 수식(修飾))

  • Kim, Se-Kwon;Lee, Eung-Ho
    • Applied Biological Chemistry
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    • v.30 no.3
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    • pp.234-241
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    • 1987
  • Conditions necessary for optimal plastein productivity from sardine protein hydrolysate using papain and pepsin were established. Sardine protein concentrate was hydrolyzed with pepsin yielding an approximate degree of hydrolysis of 77.2%. Enzyme induced plastein was optimized at: pH 6 for papain and pH 4 for pepsin; substrate concentrate, 50%(w/v) for papain and 40%(w/v) for pepsin; time of incubation, 24hr; enzyme/substrate ratio, 1 : 100(w/w). Plastein yields of 49.5% and 45.3% were found for papain and pepsin, respectively, when 10% trichloroacetic acid (TCA) was used as the precipitating agent. However, when plastein was precipitated by 50% ethanol, the yield was found to be 43.6% and 41.0% for papain and pepsin, respectively. Ethanol-precipitated plastein did not contain lipid and contained approximately 1.3% ash and 91.0% protein. In comparison, the TCA-precipitated plastein contained 74.2% protein, 0.5% lipid and 15.3% ash.

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Chemical composition of protein concentrate prepared from Yellowfin tuna Thunnus albacares roe by cook-dried process

  • Lee, Hyun Ji;Park, Sung Hwan;Yoon, In Seong;Lee, Gyoon-Woo;Kim, Yong Jung;Kim, Jin-Soo;Heu, Min Soo
    • Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.12.1-12.8
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    • 2016
  • Roe is the term used to describe fish eggs (oocytes) gathered in skeins and is one of the most valuable food products from fishery sources. Thus, means of processing are required to convert the underutilized yellowfin tuna roes (YTR) into more marketable and acceptable forms as protein concentrate. Roe protein concentrates (RPCs) were prepared by cooking condition (boil-dried concentrate, BDC and steam-dried concentrate, SDC, respectively) and un-cooking condition (freeze-dried concentrate, FDC) from yellowfin tuna roe. The yield of RPCs was in the range from 22.2 to 25.3 g/100 g of roe. RPCs contained protein (72.3-77.3 %), moisture (4.3-5.6 %), lipid (10.6-11.3 %) and ash (4.3-5.7 %) as the major constituents. The prominent amino acids of RPCs were aspartic acid, 8.7-9.2, glutamic acid, 13.1-13.2, and leucine, 8.5-8.6 g/100 g of protein. Major differences were not observed in each of the amino acid. K, S, Na, and P as minerals were the major elements in RPCs. No difference noted in sodium dodecyl sulfate polyacrylamide gel electrophoresis protein band (15-100 K) possibly representing partial hydrolysis of myosin. Therefore, RPCs from YTR could be use potential protein ingredient for human food and animal feeds.

Antioxidant activity of extracts from Blueberry (블루베리 추출물의 항산화 활성)

  • O, Jae-Yeong;Kang, Na-Lae;Kang, Seong-Woong;Song, Hyung-Yong;Kim, Hyeon-A;Hwang, Eun-Yeong;Jeon, You-Jin
    • Proceedings of the KAIS Fall Conference
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    • 2010.11b
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    • pp.744-747
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    • 2010
  • In this study, antioxidant activity of enzymatic, ethanolic and aqueous extract from Blueberry were evaluated by measuring the scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH). Enzymatic extract were prepared by enzymatic hydrolysis of Blueberry using food grade five different carbohydrases (Viscozyme, celluclast, AMG, Termarmyl, Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme, Alcalase). The ethanol extract were lower than enzymatic extracts in yield, but higher in ployphenolic contents. The 70% ethanolic extract of Blueberry exhibited better DPPH radical scavenging activity compared to those of other extracts. These results suggest that Blueberry would be a good raw materials for antioxidant.

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Studies on the $\beta$-Galactosidase from Thermophilic Bacterium - On the Production, Purification of Enzyme and the Properties of the Purified Enzyme - (고온성세균의 $\beta$-Galactosidase에 관한 연구( II )-효소의 생산, 정제 및 정제효소의 성질-)

  • 오만진;이종수;김해중;김찬조
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.15-21
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    • 1983
  • This experiment was carried out to optimize the condition for the enzyme production by selected strain in the basal medium, to purify the enzyme and to characterize the purified enzyme. The results obtained were as follows. 1. The optimal conditions for the $\beta$-galactosidase production were initial pH 7.0 and temperature $65^{\circ}C$. 2. Enzyme was induced by the addition of lactose and galactose, and it was intracellular enzyme. 3. The purified enzyme was obtained with the increased level of activity approximately 28.5 folds as compared with crude enzyme and the yield of 15.2% by means of DEAE-Cellulose column chromatography, Sephadex G-150 gel filtration 4. $\beta$-galactosidase from final step of purification showed a sing1e protein band on polyacrylamide gel disc electrophoresis. 5. The optimal temperature and pH of the purified enzyme were $65^{\circ}C$, pH 6.5 for the hydrolysis of lactose.

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Kinetic and Energetic Parameters of Carob Wastes Fermentation by Saccharomyces cerevisiae: Crabtree Effect, Ethanol Toxicity, and Invertase Repression

  • Rodrigues, B.;Peinado, J.M.;Raposo, S.;Constantino, A.;Quintas, C.;Lima-Costa, M.E.
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.837-844
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    • 2015
  • Carob waste is a useful raw material for the second-generation ethanol because 50% of its dry weight is sucrose, glucose, and fructose. To optimize the process, we have studied the influence of the initial concentration of sugars on the fermentation performance of Saccharomyces cerevisiae. With initial sugar concentrations (S0 ) of 20 g/l, the yeasts were derepressed and the ethanol produced during the exponential phase was consumed in a diauxic phase. The rate of ethanol consumption decreased with increasing S0 and disappeared at 250 g/l when the Crabtree effect was complete and almost all the sugar consumed was transformed into ethanol with a yield factor of 0.42 g/g. Sucrose hydrolysis was delayed at high S0 because of glucose repression of invertase synthesis, which was triggered at concentrations above 40 g/l. At S0 higher than 250 g/l, even when glucose had been exhausted, sucrose was hydrolyzed very slowly, probably due to an inhibition at this low water activity. Although with lower metabolic rates and longer times of fermentation, 250 g/l is considered the optimal initial concentration because it avoids the diauxic consumption of ethanol and maintains enough invertase activity to consume all the sucrose, and also avoids the inhibitions due to lower water activities at higher S0 .