• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.155-162
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• 2002

The cyclic amidohydrolase family enzymes, which include allantoinase, dihydroorotase, dihydropyrimidinase and (phenyl)hydantoinase, are metal-dependent hydrolases and play a crucial role in the metabolism of purine and pyrimidine in vivo. Each enzyme has been independently characterized, and thus well documented, but studies on the higher structural traits shared by members of this enzyme family are rare due to the lack of comparative study. Here, we report upon the expression in E. coli cells of maltose-binding protein (MBP)- and glutathione S-transferase (GST)-fused cyclic amidohydrolase family enzymes, facilitating also for both simple purification and high-level expression. Interestingly, the native quaternary structure of each enzyme was maintained even when fused with MBP and GST. We also found that in fusion proteins the favorable biochemical properties of family enzymes such as, their optimal pHs, specific activities and kinetic properties were conserved compared to the native enzymes. In addition, MBP-fused enzymes showed remarkable folding ability in-vitro. Our findings, therefore, suggest that a previously unrecognized trait of this family, namely the ability to functional fusion with some other protein but yet to retain innate properties, is conserved. We described here the structural and evolutionary implications of the properties in this family enzyme.

• 펄프종이기술
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• 제33권1호
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• pp.30-37
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• 2001

The effects of enzymatic treatment of recycled fiber were investigated to obtain the basic informations which can be used to improve the papermaking properties of recycled fiber. The recycled fibers were prepared by the repeated handsheet making and disintegrating of hardwood of hardwood and softwood kraft pulp. Novozym 342, Dinimax and Pulpzyme HC were used for enzymatic treatment. The change of fiber length distribution, freeness, contact angle and crystallinity of pulp were measured. The brightness, opacity, breaking and tear index of paper were also measured. The enzymatic treatment decreased long fiber fraction of recycled hardwood fiber, but increased long fiber fraction of recycled softwood fiber. Freeness was decreased with 0.1% enzyme and then increased again with the increase of th enzyme dosage. The improvement of flexibility of recycled fiber was obtained through the decrease of contact angle that is resulted from the decrease of crystallinity of fiber. Brightness and opacity were affected by the type of pulp and enzyme, and dosage of enzyme. Breaking length of recycled hardwood fiber was improved with enzyme treatment, but breaking length of recycled softwood fiber was decreased except for 0.01% Pulpzyme treatment. Tear index was decreased with enzymatic treatment and the lowest decrease was observed with the treatment to Pulpzyme.

• Preventive Nutrition and Food Science
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• 제2권1호
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• pp.71-76
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• 1997

Pullulanase was produced from the Klebisella pneumonias NFB_320 with the conmposition of 0.1% pullualn 1.5% yeast extract, 0.2% $K_2$HPO$_4$ and 0.02% MgSO$_4$.7$H_2O$(pH5.5). The optimum temperature for activity of the pulluanase was 3$0^{\circ}C$ and the highest yield of the enzyme was obtained after cell growth at 3$0^{\circ}C$ for 18hr, and maintained until 24hr cultivation. The pullulanase was successively purified 52.6 folds with 7.8% yield by acetone precipitation. DEAE-cellulose column chromatography and gel fitrations. The purified enzyme hydrolyzed pullulan into maltotriose exclusively. Chemical and physical properties of purified pullulanase from Klebisella pneumonias NFB-320 were examined. The optimum pH and temperature for enzyme activity were 5.0 and 6$0^{\circ}C$, respectively. The enzyme was stable between pH4 and 7, and up 5$0^{\circ}C$. The effect of mo-dification on the rate of enzyme reaction was studies with various chemicals and metal ions. The enzyme has been found to be inactivated by I$_2$ and N-bromosussinimide(NBS), which probably indicated the involve- ment of tryptophan residues in the active center of the enzyme.

• Bulletin of the Korean Chemical Society
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• 제30권1호
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• pp.57-60
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• 2009

Alcohol oxidase catalyzes the oxidation of short lines alcohol to aldehyde. In this study, alcohol oxidase from Hansenula polymorpha (HpAOD) was induced by addition of 0.5% methanol as the carbon source and purified to electrophoretic homogeneity by column chromatographies. The purified HpAOD was immobilized with DEAE-cellulose particles and its biochemical properties were compared with those of free enzyme. The substrate specificity and the optimum pH of immobilized enzyme were similar to those of free enzyme. On the other hand, the Km values of free and immobilized enzymes for ethanol were 6.66 and 14.65 mM, respectively. The optimum temperature for free enzyme was ${50^{\circ}C}$, whereas that for immobilized enzyme was ${65^{\circ}C}$. Immobilized enzyme showed high stability against long storage. Immobilized enzyme was also tested for the enzymatic determination of ethanol by the colorimetric method. We detected 1 mg/liter ethanol ($1{\times}10^{-4}$% ethanol) by 2,6- dichloroindophenol system. Therefore, the present study demonstrated that immobilized HpAOD has high substrate specificity toward ethanol and storage stability, which may be of considerable interest for alcohol biosensor and industrial application.

• 공업화학
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• 제31권4호
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• pp.352-359
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• 2020

무기 나노입자는 나노미터 크기에서 유래된 광학 및 자성 성질과 같은 물리적 특성을 활용하여 생명-의학 분야에 적극적으로 응용되어왔다. 최근에는 물리적 성질 이외에 무기 나노입자가 갖는 화학적 성질, 특히 효소와 유사한 촉매활성을 이용한 새로운 진단법들이 개발되고 있다. 효소 모사 활성의 검증에 집중하던 초기연구에서, 현재는 활성 메커니즘의 이해를 통한 적극적 활성 제어 및 치료 특성의 직접적 응용으로 연구 범위가 확장되고 있다. 본 총설에서는 효소 모사 활성을 갖는 무기 나노입자, 소위 "나노자임"의 촉매 활성 제어와 치료 및 진단 분야에서의 연구성과들에 대한 최근 동향을 정리하였다. 무기 나노입자의 효소 모사 활성은 입자의 고유한 물리적 성질과 결합되어 새로운 진단 및 치료법의 개발로 이어질 것으로 기대한다.

• 한국식품영양학회지
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• 제36권6호
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• pp.526-534
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• 2023

To improve usability of super sweet corn, extracts were prepared with hydrolytic enzyme and changes in physicochemical and antioxidant properties were analyzed. Soluble solids and reducing sugars contents were higher in all enzyme treatment groups than in the control. When enzyme treatment time increased, contents of soluble solids and reducing sugars were also increased. There was no significant difference in lightness between treatment groups, with redness showing the highest value in the control and yellowness showing the highest value in the invertase treatment group. Free sugar content in the control was the lowest. However free sugar content in the enzyme combination treatment group was increased by more than four times compared to that in the control. Contents of ascorbic acid, flavonoids and polyphenols were higher in the enzyme treatment group than in the control. In particular, the enzyme combination treatment group showed the highest content. DPPH and ABTS radical scavenging abilities were significantly higher in all enzyme treatment groups than in the control. Radical scavenging abilities of cellulase treatment group and enzyme combination treatment group showed high activity. The activity increased when enzyme treatment time increased. The combined enzyme treatment method for super sweet corn was suitable for food processing.

• 한국미생물·생명공학회지
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• 제6권1호
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• pp.33-40
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• 1978

Immobilization of alkaline protease was investigated by absorbing the enzyme on adsorbents. Alkaline protease was adsorbed on silica gel selected as a carrier to immobilize the enzyme. In this study, properties of the immobilized enzyme were compared with those of the soluble enzyme. 1) The optimum pH (10.0) of the enzyme was not changed, but the activity was increased at alkaline pH by immobilization. 2) The optimum temperature of the immobilized enzyme was shifted from 50$^{\circ}C$ to 45$^{\circ}C$, while the temperature-activity Profile became broader than those of the soluble enzyme. 3) The pH stability of the immobilized enzyme was significantely increased at pH 4.0, althouth it did not change in the neutral and alkaline pH region. 4) The heat stability of the enzyme was enhanced in the temperature range of 55$^{\circ}C$∼65$^{\circ}C$ by the immobilization. 5) The immobilized enzyme retained 40％ of its original activity after repetitive use for 6 times. 6) The enzyme stability was greately improved for a prolonged storage at 4$^{\circ}C$.

• 한국축산식품학회지
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• 제41권4호
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• pp.608-622
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• 2021

Bioactive peptides have great potentials as nutraceutical and pharmaceutical agents that can improve human health. The objectives of this research were to produce functional peptides from ovotransferrin, a major egg white protein, using single enzyme treatments, and to analyze the properties of the hydrolysates produced. Lyophilized ovotransferrin was dissolved in distilled water at 20 mg/mL, treated with protease, elastase, papain, trypsin, or α-chymotrypsin at 1% (w/v) level of substrate, and incubated for 0-24 h at the optimal temperature of each enzyme (protease 55℃, papain 37℃, elastase 25℃, trypsin 37℃, α-chymotrypsin 37℃). The hydrolysates were tested for antioxidant, metal-chelating, and antimicrobial activities. Protease, papain, trypsin, and α-chymotrypsin hydrolyzed ovotransferrin relatively well after 3 h of incubation, but it took 24 h with elastase to reach a similar degree of hydrolysis. The hydrolysates obtained after 3 h of incubation with protease, papain, trypsin, α-chymotrypsin, and after 24 h with elastase were selected as the best products to analyze their functional properties. None of the hydrolysates exhibited antioxidant properties in the oil emulsion nor antimicrobial property at 20 mg/mL concentration. However, ovotransferrin with α-chymotrypsin and with elastase had higher Fe³⁺-chelating activities (1.06±0.88%, 1.25±0.24%) than the native ovotransferrin (0.46±0.60%). Overall, the results indicated that the single-enzyme treatments of ovotransferrin were not effective to produce peptides with antioxidant, antimicrobial, or Fe³⁺-chelating activity. Further research on the effects of enzyme combinations may be needed.

• 한국식품과학회지
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• 제15권4호
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• pp.404-408
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• 1983

Malo-alcohol 발효(醱酵)에 관여하는 분열효모균(分裂酵母菌), Schizos-accharomyces japonicus var. japonicus St-3가 생성(生成)하는 malic enzyme(EC 1.1.1 40)의 몇가지 성질(性質)을 조사하였다. Malic enzyme의 생성(生成)은 배양(培養) 24시간(時間)에 최대(最大)에 이르렀고 효소반응(酵素反應)의 최적(最適) pH는 10.0, 온도(溫度)는 $25^{\circ}C$였다. 본(本) 효소(酵素)는 pH $7.0{\sim}8.4$에서 안정(安定)하였으며 $60^{\circ}C$, 10분간(分間) 열처리(熱처理)로 50% 실활(失活)되었다. $Mn^{2+}$ 첨가(添加)는 효소활성(酵素活性)을 촉진(促進)시켰으며 유기산(有機酸), 아마노산(酸) 및 ethanol의 첨가(添加)는 효소활성(酵素活性)에 아무런 영향(影響)이 없었다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.524.3-524
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• 1986

Carboxymethyl cellulase (CMCase) produced by cloned B. megaterium was found to contain 5.2% carbohydrate but no metal ion. The enzyme was isoelectric at pH 7.23 and was high is basic amino acids. The N-terminal of the enzyme was glutamic acid. The cellulolytic activity of this enzyme was extended to the small molecular substrates such as from cellotriose to cellopentaose. In additon, the enzyme showed transglycoslation activity. The pK values of the enzyme we estimated to be 4.4 and 6.7, andthat of the enzyme-substrate complex were 4.2 and 7.2, respectively. The enzyme was not affected by the treatment with iodoacetic acid, but the modification of enzyme with carbodiimide and diethyl pyrocarbonate resulted in a marked loss of the enzyme activity. These results suggest that the active site of enzyme essentially contains carboxylic and imidazole group of amino acid residues.