• Title/Summary/Keyword: fruit protease

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ISOLATION AND CHERACTERIZATION OF ACTINIDIN GENE FROM CHINESE WILD KIWI FRUIT

  • Lee, Nam-Keun;Hahm, Young-Tae
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.527-530
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    • 2000
  • A kiwi fruit ,called as the Chinese gooseberry, is originated from the Yangtze River Valley of Northern China and Zhejiang Province on the cost of Eastern China. Around 1950, a large mass production began at New Zealand with an Improved breeding. Plant origin actinidin from kiwi fruit belongs to the papain family of cysteine proteinase, which in plants includes papain from papaya, bromelain from pineapple, Cl4 protease from tomato and aleurain from barley. Actinidin is involved in the ripening-related gene family. In this study, protease gene of chinese wild kiwi fruit was isolated and characterized. 1.2kb PCR-amplified fragment was obtained from the total RNA using RT-PCR. pWACT-1 was obtained by subcloning of amplified fragment into pGEM-T Easy cloning vector and analyzed nucleotide sequence by DNA sequencing and amino acid sequence. In Result, high levels of homology between wild kiwi and New Zealand cultured-kiwi was obtained.

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Enzyme Activities of the Fruit Body of Ramaria botrytis DGUM 29001

  • Lee, Tae-Hee;Han, Yeong-Hwan
    • Mycobiology
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    • v.29 no.3
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    • pp.173-175
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    • 2001
  • The fruit body of Ramaria botrytis DGUM 29001 was used to determine enzyme activities of fruit body. The specific activity of laccase was the highest(6.5 unit/mg$\cdot$protein) and that of $\alpha$-amylase and xylanase was relatively high. However, little or no enzyme activity of $\beta$-glucosidase, CMCase, exo-$\beta$-1,4-glucanase, chitinase, lipase and protease was found.

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Protease Activity from Fruit Body of Sarcodon aspratus (능이자실체의 Protease 활성)

  • Cho, Nam-Seok;Cho, Hee-Yeon
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.4
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    • pp.58-65
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    • 2004
  • This study was performed to investigate the protease activity from fruit body of Sarcodon aspratus and its features. The specific protease activity was increased with the increasing purification steps, 2.62 times by desalting, 17 times by CMC column chromatography, 113.8 times by DEAE-Sephadex A-50 column chromatography, and 728.3 times by Sephadex G-75 column chromatography. Proteases were identified as two different enzymes having different isoelectric points at pH 4.35 (its recovery rate 8%) and pH 4.7 (its recovery rate 3.5%). Those proteases were purified by 3,025 folds and 3,257 folds in terms of specific activity. Two proteases having different isoelectric points had similar enzymatic properties. This protease was estimated to be 43,000 daltons of molecular weights by SDS-PAGE. This protease with optimum pH 4 was almost stable in the pH range of 4~7. Optimal temperature of protease activity was 40 to 50℃, and the protease activity was completely inhibited at 70℃ for 30 min.

Purification and Characterization of Metalloproteases from Pleurotus sajor-caju

  • SHIN, HYUN-HEE;HYE-SEON CHOI
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.675-678
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    • 1999
  • Fibrinolytic protease activity was detected in the fruit body of Pleurotus sajor-caju using a fibrin plate method. Two fibrinolytic activities (FPI and Ⅱ) were found at the regions of 14.5 and 86.0 kDa by using gel-filtration column chromatography. FPⅡ was identified as an alkaline protease, whereas FPⅠ was a neutral protease. Both were inhibited by phenanthrolin and EDTA, suggesting that they are metalloprotease. Inactivated enzyme activities were restored by adding Co/sup 2+/ or Zn/sup 2+/. Iodoacetate inhibited FPⅠ, but not FPⅡ. Both enzymes cleaved B/sub β/ and γ chains of the human fibrinogen. FPⅡ showed a preference to hydrophobic and bulky residues of nitroanilidine compounds as substrates, whereas FPⅠ preferred positively charged residues.

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Characteristics of Crude Protease from Fruits and Traditional Korean Fermentation Starters (과일과 전통발효제로부터 추출한 단백질분해 조효소의 비교 특성)

  • Yoo, Seon-A;Seo, Seung-Ho;Hyun, Seo-Yeon;Son, Hong-Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1461-1466
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    • 2013
  • In this study, we ascertained the characteristics of crude protease extracted from fruits (fig, kiwifruit and pineapple) as well as traditional Korean fermentation starters (bio nuruk, traditional nuruk, meju and rice koji) to determine their suitability for industrial application. Crude protease extracted from traditional Korean fermentation starters was found to have a higher optimum temperature ($70^{\circ}C$) and salt concentration (1~3%) but a lower optimum pH (3~6) value compared to the corresponding values for the protease extracted from fruits. On comparison, the total activities of protease per gram unit follow the order: bio nuruk> rice koji> traditional nuruk> fig> pineapple> meju> kiwifruit. Based on our results, we conclude that protease extracted from traditional Korean fermentation starters has potential for application in food industry, for example, as a meat tenderizer for sausage manufacturing and as a protease for cheese production.

Studies on Higher Fungi in Korea (V) -N-Terminal Amino Acid Sequence and Some Properties of Proteolytic Enzyme from Sarcodon aspratus- (한국산 고등균류에 관한 연구(제 5보) -능이 중 단백분해효소의 특성과 N-말단 아미노산배열-)

  • Eun, Jae-Soon;Yang, Jae-Hean;Lee, Tae-Kyu;Choi, Dong-Seong
    • YAKHAK HOEJI
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    • v.33 no.6
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    • pp.339-344
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    • 1989
  • The alkaline protease produced by Sarcodon aspratus(Berk) S. Ito. was purified from its fruit bodies. The enzyme was purified by using ammonium sulfate fractionation, tris-acryl CM-cellulose column chromtography and chromatofocusing. The protease migrated as one major band with a molecular weight of about 29,000 dalton on sodium dodecylsulfate-polyacrylamide gel electrophoresis. The amino acid sequence of the N-terminal residues(21) of the enzyme was determined by automated sequence analysis. The sequence was Val-Thr-Thr-Lys-Gln-Thr-Asn-Ala-Pro-Trp-Gly-Leu-Gly-Asn-Ile-Ser-Thr-Thr-Asn-Lys-Leu. Comparison of this sequence with the N-terminal sequence of the p-roteinase K from Tritirachium album showed high similarity, i. e. 57.8% identical residues. The protease displayed a relatively high stability in sodium dodecyl sulfate.

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Quality Changes of Fresh Vegetable and Fruit Juice by Various Juicers (주스제조 장치에 따른 채소 및 과일 주스의 품질 변화)

  • Choi, Moon-Hee;Kim, Min-Joo;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.29 no.3
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    • pp.145-154
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    • 2014
  • A fresh vegetable and fruit juice has become a new healthy food available for detoxification, dieting and health. This paper presents the useful information about the quality changes of fresh juice according to different juicer. Quality of fresh juice could be evaluated by several factors such as juice yield, enzyme activity, antioxidant activity, polyphenol contents, and anti-inflammatory activity. The juice yields of 12 different vegetables and fruits were compared using 6 different juicers and it was observed that the yield of slow juicer was better than that of conventional blender. Among 12 samples, the juice yield of grape is the best and the pH of the juice was in the acidic range of 3 and 4. Kiwi and grapefruit were the best in terms of protease enzyme activities by Hemoglobin units on the tyrosine basis and Spectrophotometric acid protease unit and papain units on the tyrosine basis of KFDA protocols. The total polyphenol contents were also high in kiwi and grapefruit. The antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH), superoxide dismutase (SOD), and radical scavenging assay were high in the order of kiwi, grapefruit, grape, tomato, and orange. Anti-inflammatory activities were also assay for 12 samples with 6 juicers. It can be concluded that of fresh fruit and vegetable juice provides a source of antioxidant components and enzymes with high activity. And the enzyme activities could be used as one of the quality indicator of fresh juice. Concerning the juicers used in this study, slow juicer could be recommended to prepare the fresh juice in terms of the juice quality.

The properties of Proteolytic Enzymes from the Fruit of Broussonetia Kazinoki Siebold (닥나무 열매(楮實子)에서 추출한 단백질 분해효소의 특성에 관한 연구)

  • 윤숙자;오평수;장명숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.803-806
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    • 1993
  • The properties of proteolytic enzymes from the fruit of Broussonetia Kazinoki Siebold were investigated. The protease activity of the enzymes from the fruit of Broussonetia Kazinoki Siebold was 1.6 unit. The optimum temperature and pH of the enzymes were $60^{\circ}C$ and 7.0, respectively. The enzymes were stable at pH values from 6 to 8 for 1 hr. at $37^{\circ}C$ of incubation and also retained all activity after incubation for 1 hr. at $60^{\circ}C$. The enzyme preparations showed strong activities toward hemoglobin and collagen.

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Effects of Singular Manner or Mixed Type Treatment of Proteases Isolated from Pear, Pineapple and Kiwifruit on Actomyosin Degradation (배, 파인애플 및 키위로부터 추출 분리한 단백질 분해효소의 단일 또는 혼합처리가 Actomyosin 분해에 미치는 영향)

  • 김은미;최일신;황성구
    • Food Science of Animal Resources
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    • v.23 no.3
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    • pp.193-199
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    • 2003
  • In order to investigate the meat tenderizing effects of pear, pineapple and kiwifruit, crude protease was prepared from each fruit and treated with actomyosin in a single manner or mixed type in several combination. Actomyosin was incubated with various proteases for 24 hrs under three different pH condition, and its degrading performance was evaluated by the SDS-PAGE. Pear extract showed an active degrading activity for actomyosin at pH 5.3 and 7.0. But, little actomyosin degradation was observed at pH 8.0. Actomyosin was strongly degraded by the treatment of protease from pineapple at all different pHs(5.3, 7.0 and 8.0). Kiwifruit protease extract has shown actomyosin degradation activity 1hr after treatment at pH 5.3 and pH 7.0. Meanwhile, the mixture of pear and pineapple extracts(l:l, w/w) showed much more degradation than the results of singular manner treatment at pH 5.3 and 7.0. When the pear protease was mixed with kiwifruit protease(l:l, w/w), the performance of actomyosin degradation was similar to the results of each single protease treatment. When the mixture was made of pineapple and kiwifruit extracts, actomyosin degradation was almost the same as the result of treatment of pineapple protease only. When those three proteases were mixed together(l:l:l, w/w/w), actomyosin degrading activities was in time dependent manner at pH 5.3. In summary, pear protease can be used potentially as a meat tenderizer when it was mixed with pineapple or kiwifruit rendering proper tenderization of the meat.

Inhibitory Abilities of Bacillus Isolates and Their Culture Filtrates against the Gray Mold Caused by Botrytis cinerea on Postharvest Fruit

  • Chen, Xiaomeng;Wang, Yajie;Gao, Yu;Gao, Tongguo;Zhang, Dongdong
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.425-436
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    • 2019
  • Botrytis cinerea, a major phytopathogenic fungus, has been reported to infect more than 200 crop species worldwide, and it causes massive losses in yield. The aim of this study was to evaluate the inhibitory abilities and effects of Bacillus amyloliquefaciens RS-25, Bacillus licheniformis MG-4, Bacillus subtilis Z-14, and Bacillus subtilis Pnf-4 and their culture filtrates and extracts against the gray mold caused by B. cinerea on postharvest tomato, strawberry, and grapefruit. The results revealed that the cells of Z-14, culture filtrate of RS-25, and cells of Z-14 showed the strongest biocontrol activity against the gray mold on the strawberry, grape, and tomato fruit, respectively. All the strains produced volatile organic compounds (VOCs), and the VOCs of Pnf-4 displayed the highest inhibition values. Based on headspace solid-phase microextraction in combination with gas chromatography-mass spectrometry, esters accounted for the largest percentage of the VOCs produced by RS-25, MG-4, Z-14, and Pnf-4 (36.80%, 29.58%, 30.78%, and 36.26%, respectively). All the strains showed potent cellulase and protease activities, but no chitinase activity. RS-25, Z-14, and MG-4, but not Pnf-4, grew on chrome azurol S agar, and an orange halo was formed around the colonies. All the strains showed biofilm formation, fruit colonization, and lipopeptide production, which may be the main modes of action of the antagonists against B. cinerea on the fruit. This study provides the basis for developing natural biocontrol agents against the gray mold caused by B. cinerea on postharvest fruit.