• Title/Summary/Keyword: acidic lipase

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Improved Homologous Expression of the Acidic Lipase from Aspergillus niger

  • Zhu, Si-Yuan;Xu, Yan;Yu, Xiao-Wei
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.196-205
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    • 2020
  • In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A. niger. The expression of ANL was significantly improved by the expression of the native ANL with the introns, the addition of the Kozak sequence and the optimization of the signal sequences. When the cDNA sequence of ANL fused with the glaA signal was expressed under the gpdA promoter in A. niger, no lipase activity could be detected. We then tried to improve the expression by using the full-length ANL gene containing three introns, and the lipase activity in the supernatant reached 75.80 U/ml, probably as a result of a more stable mRNA structure. The expression was further improved to 100.60 U/ml by introducing a Kozak sequence around the start codon due to a higher translation efficiency. Finally, the effects of three signal sequences including the cbhI signal, the ANL signal and the glaA signal on the lipase expression were evaluated. The transformant with the cbhI signal showed the highest lipase activity (314.67 U/ml), which was 1.90-fold and 3.13-fold higher than those with the ANL signal and the glaA signal, respectively. The acidic lipase was characterized and its highest activity was detected at pH 3.0 and a temperature of 45℃. These results provided promising strategies for the production of the acidic lipase from A. niger.

Studies on the Inhibitory Effect of Eugenia aromaticum Extract on Pancreatic Lipase

  • Kim, Seung-Kyum;Kim, Yong-Mu;Hong, Mi-Jeong;Rhee, Hae-Ik
    • Journal of Applied Biological Chemistry
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    • v.48 no.2
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    • pp.84-88
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    • 2005
  • To develop functional food and anti-obesity drug through inhibition of dietary lipid absorption, inhibitory effects of herb extracts on pancreatic lipase were investigated. Due to high yield and simplicity of isolation, lipase inhibitor (ELI) was isolated from ethyl acetate extract of Eugenia aromaticum, which showed highest inhibitory activity, and characterized for development of novel functional material. Stability of ELI at high temperature and low pH was investigated. Results showed ELI is relatively stable under thermal and acidic conditions, reversible, and noncompetitive inhibitor of pancreatic lipase.

STUDIES ON BIOLOGICALLY ACTIVE SUBSTANCES IN NON-SAPONIN FRACTION OF KOREAN RED GINSENG

  • Okuda Hiromichi;Zheng Yinan;Matsuura Yukinaga;Takaku Takeshi;Kameda Kenji
    • Proceedings of the Ginseng society Conference
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    • 1993.09a
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    • pp.110-112
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    • 1993
  • Acidic polysaccharide from Korean red ginseng was found to inhibit pancreatic lipase activity and cause reduction of plasma triglyceride level after oral administration of corn oil emulsion to rats. Thus acidic polysaccharide may reduce plasma triglyceride through its inhibitory action on pancreatic lipase and successive inhibition of intestinal absorption of fat due to reduction of lipolysis. In the course of this experiment, we found an unknown ninhydrin positive substance in Korean red ginseng. The unknown substance was identified to be arginyl-fructosyl glucose(Arg - Fru - Glc). Coment of this new compound was $5.37\%$ in Korean red ginseng powder. Sucrase and maltase activities in mucous layer of rat jejunum were found to be inhibited by Arg-Fru-Glc. Physiological significance of the new compound was discussed based on these experimental results.

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Studies on The Lypolitic Enzymes of Carum Roxburgllianum Seed Meal

  • Mahmud, Shahid;Waheed, Amran;Khanum, Razia
    • Natural Product Sciences
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    • v.10 no.6
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    • pp.302-305
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    • 2004
  • The lipase and phospholipase activities of meals of resting seeds of C. roxburghianum were studied at different temperatures, solvents and pH. Both the enzymes showed the maximum activities at $40^{\circ}C$ and in n-heptane used as solvent. However, lipase showed maximum activities at two different pH, one at pH 5 (acidic) and other at pH 8 (alkaline) whereas phospholipase showed only one pH optimum at pH 8. During the course of germination, the lipase showed an increase whereas reverse was the case with phospholipase.

Immobilization and Stability of Lipase from Mucor racemosus NRRL 3631

  • Adham, Nehad Zaki;Ahmed, Hanan Mostafa;Naim, Nadia
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.332-339
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    • 2010
  • The lipase from Mucor racemosus NRRL 3631 was partially purified by fractional precipitation using 60% ammonium sulfate, which resulted in a 8.33-fold purification. The partially purified lipase was then immobilized using different immobilization techniques: physical adsorption, ionic binding, and entrapment. Entrapment in a 4% agar proved to be the most suitable technique (82% yield), as the immobilized lipase was more stable at acidic and alkaline pHs than the free enzyme, plus 100% of the original activity was retained owing to the thermal stability of the immobilized enzyme after heat treatment for 60 min at $45^{\circ}C$. The calculated half-lives (472.5, 433.12, and 268.5 min at 50, 55, and $60^{\circ}C$, respectively) and the activation energy (9.85 kcal/mol) for the immobilized enzyme were higher than those for the free enzyme. Under the selected conditions, the immobilized enzyme had a higher $K_m$ (11.11 mM) and lower $V_{max}$ (105.26 U/mg protein) when compared with the free enzyme (8.33 mM and 125.0 U/mg protein, respectively). The operational stability of the biocatalyst was tested for both the hydrolysis of triglycerides and esterification of fatty acids with glycerol. After 4 cycles, the immobilized lipase retained approximately 50% and 80% of its original activity in the hydrolysis and esterification reactions, respectively.

Pancreatic lipase Inhibitory Compound from Apis mellifera venome

  • Kim, Jun-Ran;Kim, Shin-Duk
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.57-59
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    • 2008
  • While searching for pancreatic lipase inhibitors, the active compound was found in a methanol extract of Apis mellifera venome. The active compound was isolated by Diaion HP-20 column chromatography, thin layer chromatography and HPLC. The active compound is stable to the extreme pH and heat. There is no loss of activity both in acidic and alkaline solution in the pH range of 2 to 11 by heating for 15 minutes at $90^{\circ}C$. The rf value of the compound was 0.51 at TLC with butanol : methanol: water (4:1:2) solvent system. The molecular weight of the compound was determined to be 293 by EI-MS.

In vitro stability evaluation of coated lipase

  • Liu, Lu Jie;Zhu, Jia;Wang, Bin;Cheng, Chu;Du, Yong Jie;Wang, Min Qi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.192-197
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    • 2017
  • Objective: The study was conducted to evaluate the stability of commercial coated lipase (CT-LIP) in vitro. Methods: The capsules were tested under different conditions with a range of temperature, pH, dry heat treatment and steaming treatment, simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) in this work, respectively. Free lipase (uncoated lipase, UC-LIP) was the control group. Lipase relative activities measured in various treatments were used as a reference frame to characterize the stability. Results: The lipase activities were decreased with increasing temperatures (p<0.05), and there was a markedly decline (p<0.01) in lipase comparative activities of UC-LIP at $80^{\circ}C$ compared with CT-LIP group. Higher relative activities of lipase were observed in CT-LIP group compared with the free one under acidic ambient (pH 3 to 7) and an alkaline medium (pH 8 to 12). Residual lipase activities of CT-LIP group were increased (p<0.05) by 5.67% and 35.60% in dry heat and hydrothermal treatments, respectively. The lipase relative activity profile of CT-LIP was raised at first and dropped subsequently (p<0.05) compared with constantly reduced tendency of UC-LIP exposed to both SGF and SIF. Conclusion: The results suggest that the CT-LIP possesses relatively higher stability in comparison with the UC-LIP in vitro. The CT-LIP could retain the potential property to provide sustained release of lipase and thus improved its bioavailability in the gastrointestinal tract.

Comparison of Feed Efficiency Between Rotifers Enriched Lipid-contents to Enrichment and Enhanced Digestive Enzymes Activity to Starch (영양강화 Rotifer와 효소활성 향상 Rotifer의 먹이효율 비교)

  • Kwon, O-Nam;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.22 no.1
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    • pp.105-111
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    • 2009
  • In this study, we carried out an experiment for estimation the larval digestibility in aspects which digestive enzymatic activities and nutrition of the rotifers, Brachionus rotundiformis. Thus we enhanced the digestive enzymatic activity through the addition of starch for the increase of digestibility of rotifer (starch-rotifer), and compared with the feed efficiency through rearing of the olive flounder, Paralichthys olivaceus used rotifer lipid-enriched with Algamac $2000^{(R)}$ (CE-rotifer). The digestive enzyme activities (except for TG-lipase), total protein contents, total essential amino acid, essential amino acids (methionin and phenylalanine) of starch-rotifer (the rotifer used a starch as additive, and enriched not) was assayed significantly higher than CE-rotifer (P<0.05). And total lipid, lipid classes (except for sterol) and fatty acids as DHA and EPA showed higher in CE-rotifer than starch-rotifer (P<0.05). But, sterol contents and ST/TG ratio were shown significantly higher in starch-rotifer (P<0.05). The flounder larvae supplied the two rotifers showed standard length and body weight that not significantly differed with ranges $3.72{\sim}3.79\;mm$ and $32.9{\sim}37.8\;mg$/larva on 6 days after hatching (DAH), respectively (P>0.05). However, these of 12 DAH showed the values of significantly higher to $5.94{\pm}0.249\;mm$, $144.0{\pm}23.86\;mg$/larva and $26.2{\pm}12.13%$ in standard length, body weight and survival in CE-flounder than that of starch-flounder (P<0.05). The hydrolytic enzymatic activities of flounder larvae severally supplied the two rotifers showed the significantly higher activities in acidic -amylase, neutral -amylase, TG-lipase, lysozyme and acidic phosphatase in starch-flounder on 5 DAH (P<0.05). But neutral $\alpha$-amylase, three proteases and two phosphatases of CE-flounder on 11 DAH showed the significantly higher activities than that of starch-flounder (P<0.05). Therefore, for the flounder, Paralichthys olivaceus larvae just depleted yolk was more beneficial to supply the feed, rotifer, enhanced the digestibility than to supply the feed lipid-enriched for aspect of larval digestibility up to 6 DAH, thereafter nutrition of absorption due to the development of digestive organs suggested that enrichment effect appeared with larval somatic growth. Consequently, investigation more detailed about the larval digestive physiological and nutritional requirement variations after 6 DAH will be necessary, thereafter.

Characteristics of Traditional Mejus of Nation-Wide Collection (전국적으로 수집한 전통식 메주의 특성 조사)

  • 유진영;김현규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.2
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    • pp.259-267
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    • 1998
  • Typical characteristics of Mejus must be understood to get the basic data for setting up mass production system of traditional fermented soybean products. One hundred and twenty one Mejus were collected from various places and analyed. Most of shapes were rectangular and some were spherical, conical, cylindrical and doughnut types. The weight of Mejus was 0.4~4.2kg. Chemical analysis showed: moisture content, 9.73~58.22% ; pH, 4.95~8.15; acidity, 0.6~3.8% ; soluble protein content, 4.45~12.31%; soluble sugar content, 0.82~10.95%. Enzyme assay showed: $\alpha$-amylase activity, 5.0~874.2 units/g; $\beta$-amylase activity, 0.02~27.74units/g; acidic protease activity, 31.3~225.1unts/g; lipase activity 1.0~53.0units/g. Total viable cells were 3.72$\times$107~1.35$\times$1010cfu/g, and yeast and mold count 6.46$\times$104~8.91$\times$106cfu/g. respectively. $\alpha$-Amylase activity of a traditional Meju from Incheon showed the highest activity of 732.8 units/g(interior section) and 823.2units/g (exterior section). $\beta$-Amylase activity was the highest{3.57 units/g (interior sectin) and 4.25units/g (exterior section)} in Meju from Chunbuk. Acidic protease activity was the highest in sample from Seoul, whereas traditional Meju from Kyongnam showed the highest activity of 21.5units/g(interior section) and 37.5units/g(exterior sectin).

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Physico-chemical and Microbiological Changes of Traditional Meju during Fermentation in Kangweondo Area (강원도 지방의 재래식 메주 발효중 이화학적 특성 및 미생물의 변화)

  • Yoo, Jin-Young;Kim, Hyeon-Gyu;Kim, Wang-June
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.908-915
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    • 1998
  • By using Korean native soybean, traditional meju was prepared in Chuncheon, Kangweondo according to the traditional process. Analysis of physico-chemical, enzymatic and microbiological changes during meju fermentation were carried out in order to obtain a basic information for industrial scale production of meju. The enviroments for natural meju fermentation were $10{\sim}15^{\circ}C$ and $60{\sim}70%{\;}RH$. Moisture content decreased from 59% to 11% (exterior section) and 19% (interior section). the pH of meju rapidly increased up to 8.5 at $33^{rd}{\;}day$ of fermentation and thereafter decreased down to 7.9 at $70^{th}{\;}day$ of fermentation. Souble protein content was 1.47% at initial stage and increased up to $6.31{\sim}7.34%$ at $33^{rd}{\;}day$ of fermentation. Amino nitrogen content was $460{\sim}770{\;}mg%$ at $70^{th}{\;}day$ of fermentation. the color of meju became gradually black and decreased in redness and yellowness. During the process, protease and lipase seemed to play an important role in the digestion of soy protein and fat. Acidic protease activity increased up to $135.9{\sim}152.4{\;}unit/g$ at $33^{rd}{\;}day$ of fermentation and were $181.3{\sim}272.6{\;}unit/g$ at $70^{th}{\;}day$ of fermentation. Lipase activity increased up to 6 unit/g (interior section) and 15 unit/g (exterior section) at $70^{th}{\;}day$ of fermentation. the viable cell count of meju was at the level of $10^8{\;}CFU/g$ during the overall fermentation period. Aerobic halophilic count was $1.51{\times}10^7{\;}CFU/g$ at initial stage and maintained $10^8{\;}CFU/g$ level during the process. Initial anaerobic cell count was $2.0^9{\times}10^4{\;}CFU/g$ and increased up to $10^5{\;}CFU/g$ level at 47 days. Yeast and mold counts were $10^4{\sim}10^5{\;}CFU/g$ for the fermentation period.

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