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中国精品科技期刊2020
谌新然, 黄永生, 田康明, 金鹏, 董自星, 刘晓光, 路福平, 王正祥. 重组聚半乳糖醛酸酶PGB的生化特征及其酶解产物活性分析[J]. 食品工业科技, 2017, (23): 119-123. DOI: 10.13386/j.issn1002-0306.2017.23.024
引用本文: 谌新然, 黄永生, 田康明, 金鹏, 董自星, 刘晓光, 路福平, 王正祥. 重组聚半乳糖醛酸酶PGB的生化特征及其酶解产物活性分析[J]. 食品工业科技, 2017, (23): 119-123. DOI: 10.13386/j.issn1002-0306.2017.23.024
CHEN Xin-ran, HUANG Yong-sheng, TIAN Kang-ming, JIN Peng, DONG Zi-xing, LIU Xiao-guang, LU Fu-ping, WANG Zheng-xiang. Biochemical characterization of the recombinant polygalacturonase PGB and the activity analysis of its enzymatic hydrolysis product[J]. Science and Technology of Food Industry, 2017, (23): 119-123. DOI: 10.13386/j.issn1002-0306.2017.23.024
Citation: CHEN Xin-ran, HUANG Yong-sheng, TIAN Kang-ming, JIN Peng, DONG Zi-xing, LIU Xiao-guang, LU Fu-ping, WANG Zheng-xiang. Biochemical characterization of the recombinant polygalacturonase PGB and the activity analysis of its enzymatic hydrolysis product[J]. Science and Technology of Food Industry, 2017, (23): 119-123. DOI: 10.13386/j.issn1002-0306.2017.23.024

重组聚半乳糖醛酸酶PGB的生化特征及其酶解产物活性分析

Biochemical characterization of the recombinant polygalacturonase PGB and the activity analysis of its enzymatic hydrolysis product

  • 摘要: 解析重组聚半乳糖醛酸酶PGB的酶学性质,研究PGB制备的果胶水解物的基本成分和生物学活性。通过摇瓶发酵制备聚半乳糖醛酸酶酶液,研究其基本酶学性质。再在最优作用条件下对果胶进行水解,并考察水解产物的组成、抗菌活性和抗氧化活性。在摇瓶发酵水平,黑曲霉聚半乳糖醛酸酶PGB的酶活达到10790 U/m L;该酶的最适作用温度和p H分别为55℃和5.0;在40℃或p H4.06.0条件下具有较好稳定性。Mg2+和Cu2+对其活性有促进作用,K+、Ca2+、Zn2+、Mn2+、Co2+、Fe3+和Fe2+等对其活性均有不同程度的抑制作用;它对低酯果胶的Km和Vmax分别是0.756 mg/m L和9.225 mg/(m L·min)。高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD)的分析表明,PGB对果胶的酶解产物是半乳糖醛酸单体和聚合度24为主的寡聚半乳糖醛酸,它对金黄色葡萄球菌的生长有显著抑制作用,其最小抑菌浓度和抗氧化活性分别为0.64 mg/m L和6.125 U/m L。 

     

    Abstract: To analyze the enzymatic properties of recombinant polygalacturonase PGB and to study the compositions and biological activities of pectin hydrolysate prepared by PGB.Polygalacturonase was prepared by shake flask fermentation and its enzymatic properties were investigated.Pectin was hydrolyzed by PGB under optimum conditions, followed by investigation of its compositions, and antioxidant and antibacterial activities. At the shake flask fermentation level, the activity of PGB reached10790 U/m L.Its optimum temperature and p H was 55 ℃ and 5.0, respectively.And it was quite stable at 40 ℃ or p H4.0 6.0.Mg2+and Cu2+enhanced its activity, while K+, Ca2+, Zn2+, Mn2+, Co2+, Fe3 +and Fe2+had different inhibitory effects on its activity.The Kmand Vmaxof PGB toward low-ester pectin was determined to be 0.756 mg/m L and 9.225 mg/ ( m L·min) . After hydrolyzed by PGB, pectin hydrolysate was identified as galacturonic acid and oligogalacturonic acid with a polymerization degree of 2 to 4 by high performance anion exchange chromatography-pulsed amperometric detector ( HPAEC-PAD) . This oligomer significantly inhibited the growth of Staphylococcus aureus, and its minimum inhibitory concentration and antioxidant activity was 0.64 mg/m L and 6.125 U/m L, respectively.

     

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