• Journal of Microbiology and Biotechnology
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• 제29권6호
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• pp.944-951
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• 2019

Lipases are industrial enzymes that catalyze both triglyceride hydrolysis and ester synthesis. The overexpression of lipase genes is considered one of the best approaches to increase the enzymatic production for industrial applications. Subfamily I.2. lipases require a chaperone or foldase in order to become a fully-activated enzyme. The goal of this research was to isolate, clone, and co-express genes that encode lipase and foldase from Burkholderia territorii GP3, a lipolytic bacterial isolate obtained from Mount Papandayan soil via growth on Soil Extract Rhodamine Agar. Genes that encode for lipase (lipBT) and foldase (lifBT) were successfully cloned from this isolate and co-expressed in the E. coli BL21 background. The highest expression was shown in E. coli BL21 (DE3) pLysS, using pET15b expression vector. LipBT was particulary unique as it showed highest activity with optimum temperature of $80^{\circ}C$ at pH 11.0. The optimum substrate for enzyme activity was $C_{10}$, which is highly stable in methanol solvent. The enzyme was strongly activated by $Ca^{2+}$, $Mg^{2+}$, and strongly inhibited by $Fe^{2+}$ and $Zn^{2+}$. In addition, the enzyme was stable and compatible in non-ionic surfactant, and was strongly incompatible in ionic surfactant.

• 한국의류학회지
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• 제37권8호
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• pp.1107-1116
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• 2013

This study examined the use of cutinase and lipase to process cotton/polyester blend fabric. Optimum treatment conditions for cutinase and lipase were investigated for cotton/polyester blend fabric. The properties of enzyme-treated fabrics were evaluated and compared in optimal treatment conditions. In addition, the possibility to provide an enzymatic finishing on blend fabrics using mixed enzymes in a two-step process were studied. The weight loss of cotton/polyester blend fabrics with Triton X-100 was 0.8% and the dyeing property of blend fabrics with calcium chloride increased by a factor of 1.2. The use of two enzymes in combination with cutinase and lipase in the presence of auxiliaries resulted in a cotton/polyester blend fabric weight loss of 0.8%. In addition, the dyeing properties of cotton/polyester blend fabrics improved by a factor of 1.5 and the moisture regain of cotton/polyester blend fabrics improved by a factor of 1.16. However, no marked loss was observed in tensile strength. The surface morphology of cotton/polyester blend fabrics is modified through a two-enzyme treatment. The treatment of cotton/polyester blend fabrics with cutinase and lipase maintains cotton strength and improves the moisture regain of polyester fabrics.

• 한국식품과학회지
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• 제8권3호
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• pp.141-146
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• 1976

1) 조효소액(粗酵素液)을 유안염석(硫安鹽析)(0.5포화도(飽和度)) Sephadex G 25에 의(依)한 제염(除鹽), CM Cellulose column chromatography, Sephadex G 25에 의(依)한 농축(濃縮), Sephadex G 75 gel filtration에 의(依)하여 Specific activity 126.5/mg protein, 원활성(原活性)의 약(約) 45배(倍), 수율(收率) 4.2%의 정제효소(精製酵素)를 얻었다. 2) 정제효소(精製酵素)를 Acrylamide gel disc electrophoresis에 의(依)하여 분리(分離)시킨 결과(結果) 하나의 주(主)된 band와 그 양측(兩側)에 2개(個)의 희미한 band가 나타나 있으므로 하나의 순수(純粹)한 단백질(蛋白質)이라고는 할 수 없으나 조효소액(粗酵素液)에 비(比)하여 상당(相當)히 정제(精製)되었다.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.216-225
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• 2020

An esterase gene, estA1, was cloned from Alteromonas sp. 39-G1 isolated from the Beaufort Sea. The gene is composed of 1,140 nucleotides and codes for a 41,190 Da protein containing 379 amino acids. As a result of a BLAST search, the protein sequence of esterase EstA1 was found to be identical to Alteromonas sp. esterase (GenBank: PHS53692). As far as we know, no research on this enzyme has yet been conducted. Phylogenetic analysis showed that esterase EstA1 was a member of the bacterial lipolytic enzyme family IV (hormone sensitive lipases). Two deletion mutants (Δ20 and Δ54) of the esterase EstA1 were produced in Escherichia coli BL21 (DE3) cells with part of the N-terminal of the protein removed and His-tag attached to the C-terminal. These enzymes exhibited the highest activity toward p-nitrophenyl (pNP) acetate (C₂) and had little or no activity towards pNP-esters with acyl chains longer than C₆. Their optimum temperature and pH of the catalytic activity were 45℃ and pH 8.0, respectively. As the NaCl concentration increased, their enzyme activities continued to increase and the highest enzyme activities were measured in 5 M NaCl. These enzymes were found to be stable for up to 8 h in the concentration of 3-5 M NaCl. Moreover, they have been found to be stable for various metal ions, detergents and organic solvents. These salt-tolerant and chemical-resistant properties suggest that the enzyme esterase EstA1 is both academically and industrially useful.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1260-1268
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• 2014

Screening of a gene library from Paenibacillus sp. PBS-2 generated in Escherichia coli led to the identification of a clone with lipolytic activity. Sequence analysis showed an open reading frame encoding a polypeptide of 378 amino acid residues with a predicted molecular mass of 42 kDa. The esterase displayed 69% and 42% identity with the putative ${\beta}$-lactamases from Paenibacillus sp. JDR-2 and Clostridium sp. BNL1100, respectively. The esterase contained a Ser-x-x-Lys motif that is conserved among all ${\beta}$-lactamases found to date. The protein PBS-2 was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme is a serine protein and was active against p-nitrophenyl esters of $C_2$, $C_4$, $C_8$, and $C_{10}$. The optimum pH and temperature for enzyme activity were pH 9.0 and $30^{\circ}C$, respectively. Relative activity of 55% remained at up to $5^{\circ}C$ with an activation energy of 5.84 kcal/mol, which indicates that the enzyme is cold-adapted. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, and $Hg^{2+}$ ions. As expected for a serine esterase, activity was inhibited by phenylmethylsulfonyl fluoride. The enzyme was remarkably active and stable in the presence of commercial detergents and organic solvents. This cold-adapted esterase has potential as a biocatalyst and detergent additive for use at low temperatures.

• The Korean Journal of Ecology
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• 제18권3호
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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.

• 한국식품영양학회지
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• 제9권4호
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• pp.392-397
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• 1996

저식염 멸치젓을 속성으로 제조하기 위하여 Asp. oryzae와 Bacillus sp.로 만든 zdh지를 첨가하고 숙성중 미생물상 변화와 효소활성을 비교 검토하였다. 젓갈 숙성 중 미생물상은 단백질분해균과 혐기성 균수는 숙성 40일경, 호기성 균수는 20일경에 많았다. 단백질분해균과 지방질분해균, 호기성균, 혐기성 균수는 Bacillus sp. 코오지 첨가구에서 높았다. 젓갈 숙성중 단백질 가수분해효소 활성은 숙성 20일 경에, 지방질 가수분해효소는 숙성 30일 경에 높았다가 점진적으로 감소하였으며, 단백질 가수분해효소는 Asp. oryzae 코오지 첨가구가, 지방질 가수분해효소는 Bacillus sp. 코오지 첨가구에서 높았다.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.2024-2033
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• 2015

A gene encoding lipolytic enzyme, lip7-3, was isolated from Bacillus sp. W130-35 isolated from a tidal mud flat. The gene encoded a protein of 215 amino acids with a signal peptide composed of 34 amino acid residues. Lip7-3 belonged to the family I.4 lipase and showed its maximal activity at pH 9.0 and 60℃. Its activity increased in the presence of 30% methanol and, remarkably, increased as well to 154.6% in the presence of Ca²⁺. Lip7-3 preferred p-nitrophenyl octanoate (C8) as a substrate and exhibited broad specificity for short- to long- chain fatty acid esters. Additionally, Lip7-3 showed a low degree of enantioselectivity for an S-enantiomer (e.g., (S)-methyl-3-hydroxy-2-methylpropionate). It efficiently hydrolyzed glyceryl tributyrate, but did not hydrolyze glyceryl trioleate, fish oil, or olive oil. Its substrate specificity and activation by the solvent might offer a merit to the biotechnological enzyme applications like transesterification in the production of biodiesel.

• Journal of Dairy Science and Biotechnology
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• 제36권1호
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• pp.49-71
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• 2018

Heat treatment is the most popular processing technique in the dairy industry. Its main purpose is to destroy the pathogenic and spoilage bacteria in order to ensure that the milk is safe throughout its shelf life. The protease and lipase that are present in raw milk might reduce the quality of milk. Plasmin and protease, which are produced by psychrotrophic bacteria, are recognized as the main causes of the deterioration in milk flavor and taste during storage. The enzymes in raw milk can be inactivated by heat treatment. However, the temperature of inactivation varies according to the type of enzyme. For example, some Pseudomonas spp. produce heat-resistant proteolytic and lipolytic enzymes that may not be fully inactivated by the low temperature and long time (LTLT) treatment. These types of enzymes are inhibited only by the high temperature and short time (HTST) or ultra-high temperature (UHT) treatment of milk.

• 한국축산식품학회지
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• 제34권1호
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• pp.40-48
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• 2014

In this study, the effects of two different commercial starter culture mixes and processing methodologies (traditional and heat process) on the lipolytic changes of fermented sausages manufactured with turkey meat were evaluated during processing stages and storage. Free fatty acid (FFA) value increased with fermentation and during storage over 120 d in all fermented sausage groups produced with both processing methodologies (p<0.05). After drying stage, free fatty acid values of traditional style and heat processed fermented sausages were between 10.54-13.01% and 6.56-8.49%, respectively. Thiobarbituric acid (TBA) values of traditionally processed fermented sausages were between $0.220-0.450mg{\cdot}kg^{-1}$, and TBA values of heat processed fermented sausages were in a range of $0.405-0.795mg{\cdot}kg^{-1}$. Oleic and linoleic acids were predominant fatty acids in all fermented sausages. It was seen that fermented sausage groups produced with starter culture had lower TBA and FFA values in comparison with the control groups, and heat application inhibited the lipase enzyme activity and had an improving effect on lipid oxidation. As a result of these effects, heat processed fermented sausages had lower FFA and higher TBA values than the traditionally processed groups.