目的:研究黑线鳕鱼(Melanogrammus aeglefinus)皮胶原蛋白胰蛋白酶酶解多肽的抗氧化、抑制光损伤作用,提高鱼皮的再利用价值。方法:利用单因素试验和响应面试验设计,探讨黑线鳕鱼皮胶原蛋白的最佳酶解工艺。将制得的酶解多肽通过基质辅...目的:研究黑线鳕鱼(Melanogrammus aeglefinus)皮胶原蛋白胰蛋白酶酶解多肽的抗氧化、抑制光损伤作用,提高鱼皮的再利用价值。方法:利用单因素试验和响应面试验设计,探讨黑线鳕鱼皮胶原蛋白的最佳酶解工艺。将制得的酶解多肽通过基质辅助激光解离飞行时间串联质谱仪(matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry,MALDI-TOF MS/MS)测定多肽序列。利用Discovery Studio中CDOCKER模块将所得的优势肽与抗氧化相关的Kelch样环氧氯丙烷相关蛋白-1(Keap1)进行分子对接。用不同质量浓度的多肽预处理人表皮角质形成细胞(Ha Ca T)后,使用中波紫外线诱导HaCaT细胞构建光损伤模型,通过超氧化歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、丙二醛(malondialdehyde,MDA)含量及细胞凋亡与周期来研究酶解多肽抑制光损伤的能力。结果:黑线鳕鱼皮胶原蛋白在胰蛋白酶质量分数1.23%、酶解温度52.95℃、酶解时间1.75 h条件下,水解度为78.33%。MALDI-TOF MS/MS测定出酶解液中的优势肽序列为V-IFFVTMGTP-R、L-SIVPV-R和L-MHT-T。通过分子对接发现V-IFFVTMGTP-R与Keap1结合最稳定,从而预测其具有抗氧化的作用。用250 mg/m L酶解液处理光损伤的HaCaT细胞,其SOD、GSHPx活性较模型组极显著提高(P<0.01),MDA含量极显著降低(P<0.01)。结论:黑线鳕鱼皮胶原蛋白胰蛋白酶酶解多肽具有抗氧化活性,对光损伤具有一定的抑制作用。展开更多
[Objective] The paper was to identify strain HN-1 against banana wilt disease and to determine its antagonism. [Method] The strain HN-1 was ob- tained from the soil in fields heavily infected by Fusarium oxysporum f. ...[Objective] The paper was to identify strain HN-1 against banana wilt disease and to determine its antagonism. [Method] The strain HN-1 was ob- tained from the soil in fields heavily infected by Fusarium oxysporum f. sp. cubense （FOC）. Antagonism of the strain against F. oxysporum was tested via dual-cul- ture and inhibition test on spore germination. [Result] HN-1 effectively inhibited mycelial growth and spore germination of F. oxysporum, Strain HN-1 was identi- fied as BrevibaciUus brevis according to its characteristics in morphology, physiology and biochemistry and its 16S rDNA sequence. The strain showed high inhibition effect on 15 species of fungal pathogens in the dual-culture trials with fungal pathogens. [ Conclusion] The study provides theoretical basis for application of strain HN-1 in agricultural fields.展开更多
文摘目的:研究黑线鳕鱼(Melanogrammus aeglefinus)皮胶原蛋白胰蛋白酶酶解多肽的抗氧化、抑制光损伤作用,提高鱼皮的再利用价值。方法:利用单因素试验和响应面试验设计,探讨黑线鳕鱼皮胶原蛋白的最佳酶解工艺。将制得的酶解多肽通过基质辅助激光解离飞行时间串联质谱仪(matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry,MALDI-TOF MS/MS)测定多肽序列。利用Discovery Studio中CDOCKER模块将所得的优势肽与抗氧化相关的Kelch样环氧氯丙烷相关蛋白-1(Keap1)进行分子对接。用不同质量浓度的多肽预处理人表皮角质形成细胞(Ha Ca T)后,使用中波紫外线诱导HaCaT细胞构建光损伤模型,通过超氧化歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、丙二醛(malondialdehyde,MDA)含量及细胞凋亡与周期来研究酶解多肽抑制光损伤的能力。结果:黑线鳕鱼皮胶原蛋白在胰蛋白酶质量分数1.23%、酶解温度52.95℃、酶解时间1.75 h条件下,水解度为78.33%。MALDI-TOF MS/MS测定出酶解液中的优势肽序列为V-IFFVTMGTP-R、L-SIVPV-R和L-MHT-T。通过分子对接发现V-IFFVTMGTP-R与Keap1结合最稳定,从而预测其具有抗氧化的作用。用250 mg/m L酶解液处理光损伤的HaCaT细胞,其SOD、GSHPx活性较模型组极显著提高(P<0.01),MDA含量极显著降低(P<0.01)。结论:黑线鳕鱼皮胶原蛋白胰蛋白酶酶解多肽具有抗氧化活性,对光损伤具有一定的抑制作用。
基金Supported by Special Funds for Construction of Modern Agricultural Technology System(nycytx-24)Fundamental Research Special Project for Central Public-interest Scientific Institutes(ITBB130502)
文摘[Objective] The paper was to identify strain HN-1 against banana wilt disease and to determine its antagonism. [Method] The strain HN-1 was ob- tained from the soil in fields heavily infected by Fusarium oxysporum f. sp. cubense （FOC）. Antagonism of the strain against F. oxysporum was tested via dual-cul- ture and inhibition test on spore germination. [Result] HN-1 effectively inhibited mycelial growth and spore germination of F. oxysporum, Strain HN-1 was identi- fied as BrevibaciUus brevis according to its characteristics in morphology, physiology and biochemistry and its 16S rDNA sequence. The strain showed high inhibition effect on 15 species of fungal pathogens in the dual-culture trials with fungal pathogens. [ Conclusion] The study provides theoretical basis for application of strain HN-1 in agricultural fields.
