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【目的】研究影响油菜籽饼粕中硫代葡萄糖苷在黑芥子酶作用下定向转化为异硫氰酸酯的因素,确定最佳酶解条件,探索外加黑芥子酶对硫代葡萄糖苷降解产物组分和含量的影响,为油菜籽饼粕的综合利用提供基础。【方法】测定4种反应体系中黑芥子酶活性,确定酶解反应缓冲体系。以油菜籽饼粕中异硫氰酸酯总量为指标,单因素结合正交设计研究不同料液比、时间、温度、pH、抗坏血酸添加量对外加黑芥子酶降解油菜籽饼粕中硫代葡萄糖苷转化为异硫氰酸酯的影响,优化工艺参数,获得最佳酶解条件。采用二氯甲烷为提取剂,真空浓缩得到油状降解产物,利用气相色谱-质谱（GC-MS）联用鉴定外加黑芥子酶和内源黑芥子酶酶解产物成分,分析产物相对含量。【结果】4种缓冲体系中,在柠檬酸-磷酸盐缓冲液中测得黑芥子酶活性最高,在相同条件下,外加黑芥子酶的活性显著高于内源黑芥子酶活性;外源黑芥子酶添加量大于油菜籽饼粕量的20%,异硫氰酸酯生成量无显著增加,选择外源黑芥子酶添加量为油菜籽饼粕量的20%;正交优化得到最佳酶解条件为：料液比1﹕15,pH为6.0,反应温度为50℃,抗坏血酸添加量为0.005 mg•g-1,反应时间为1 h,异硫氰酸酯含量达到1.799 mg•g-1,是油菜籽饼粕内源黑芥子酶降解硫代葡萄糖苷生成异硫氰酸酯产物总量的7.6倍。外加黑芥子酶生成异硫氰酸酯含量显著高于内源黑芥子酶降解硫苷生成异硫氰酸酯含量,但两者异硫氰酸酯含量随反应条件变化趋势一致。GC-MS鉴定外加黑芥子酶有7种酶解产物,相对含量在产物的67%以上,含有1种腈类,2种噁唑烷酮和4种异硫氰酸酯。产物中异硫氰酸酯为烯丙基异硫氰酸酯、丁烯基异硫氰酸酯、环戊基异硫氰酸酯和苯乙基异硫氰酸酯;其中丁烯基异硫氰酸酯的含量最高,占酶解产物总量的26.77%。腈类产物为苯丙腈;2种噁唑烷酮为致甲状腺肿素和2,1-苯丙噁唑烷酮。内源黑芥子酶降解硫代葡萄糖苷的产物中检测到4种产物且含量相对较少。【结论】外加黑芥子酶能有效增加油菜籽饼粕中异硫氰酸酯生成量,确定了油菜籽饼粕中硫代葡萄糖苷的最佳降解条件。鉴定出降解产物的主要成分,异硫氰酸酯有4种,其中丁烯基异硫氰酸酯含量最高。 相似文献
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油菜饼中硫代葡萄糖苷含量的遗传及其低含量品种的选育 总被引：1，自引：0，他引：1
傅廷栋 《华中农业大学学报》1982,(3)
一芸苔属植物中的硫代葡萄糖苷主要有5种(A.Gland,1980),即:黑芥子硫苷酸钾(Sinigrin),葡萄糖苷(gluconapin),芸苔葡萄糖苷(glucobrassica),甲状腺素(Progoitrin)和napoleifierin。这五种硫代葡萄糖苷均属烯烃类硫代葡萄糖苷。硫代葡萄糖苷本身无毒,但它在一组芥子酶的水解作用下,产生恶唑烷硫铜(OZT,主要是甲状腺素和napoleiferin的裂解产物)、异硫氰酸盐(主要是黑芥子硫苷硫钾、葡萄糖苷和芸苔葡萄糖苷的裂解产物)和氰 相似文献
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采用甲醇法提取西兰花中硫代葡萄糖苷,利用重量法测定其中硫代葡萄糖苷的含量,得出硫代葡萄糖苷最佳提取条件,并对硫代葡萄糖苷提取液进行酸水解,通过抑菌圈实验,分析了西兰花中硫代葡萄糖苷水解产物异硫氰酸酯类的抑菌性。其结果表明,采用80％甲醇溶液、料液比为10：40（w／v）、80℃为提取的最佳条件。西兰花花蕾中硫代葡萄糖苷的含量最高,达到39．90μmol／g;花茎中为18．28μmol／g：叶中为11．86μmol／g。实验结果表明异硫氰酸酯类对金黄色葡萄球菌（Staphylococcus aureus）、白葡萄球菌（White staphylococcus）、枯草芽孢杆菌（Bacillus subtilis）大肠杆菌（Escherichia coli）的抑菌作用均较明显。 相似文献
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硫代葡萄糖苷是一种含硫的阴离子亲水性植物次生代谢产物,广泛存在于芸苔属植物的籽实或粗饲料中,其含量和组成由于植物种类、栽培方法和气候环境等条件的不同而变化很大。菜籽饼粕中含有大量的硫代葡萄糖苷,从而影响了饲料的适口性,并且摄入过多容易引起动物中毒,动物生长缓慢,生产性能降低。文章对硫代葡萄糖苷的分布、抗营养作用及在动物体内的代谢进行了综述。 相似文献
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十字花科蔬菜硫代葡萄糖苷含量比较 总被引：6，自引：0，他引：6
本试验采用硫酸根离子沉淀法对十字花科蔬菜中白菜类、甘蓝类、芥菜类、萝卜类蔬菜的硫代葡萄糖苷含量进行了测定.结果表明,甘蓝类的硫代葡萄糖苷的含量最高,是白菜类和芥菜类的10倍多,是萝卜类肉质根中的15倍多.在甘蓝类蔬菜中,普通结球甘蓝的硫代葡萄糖苷含量较高.在白菜类蔬菜中,大白菜含有较高的硫代葡萄糖苷,普通油菜和菜薹的含量则稍低.大叶芥菜的硫代葡萄糖苷含量界于大白菜和普通油菜含量之间.在萝卜的肉质根中,北京水萝卜和白萝卜硫代葡萄糖苷较高,菜薹的叶片中硫代葡萄糖苷的含量比其嫩茎中高出1倍多. 相似文献
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《浙江大学学报(农业与生命科学版)》2020,(3)
5-氨基乙酰丙酸(5-aminolevulinic acid, ALA)是一种可以影响植物碳固定和营养物质同化等生理生化过程的植物生长调节剂。本文研究了ALA对幼苗期甘蓝型油菜硫代谢和硫代葡萄糖苷生物合成的影响。用含有0、0.5、1.0、5.0、10.0 mg/L ALA的霍格兰培养液处理油菜幼苗28 d后,分析ALA对其硫醇和硫代葡萄糖苷含量以及相关基因转录组的影响。结果表明:低质量浓度的ALA处理增加了半胱氨酸、硫代葡萄糖苷以及总可溶性硫醇的含量,同时上调了调节硫摄取和代谢相关基因的表达,如Bn SULTR1.1、Bn SULTR2.2和Bn APK1。由于硫代谢的加速以及甲硫氨酸的积累,ALA还提高了硫代葡萄糖苷的含量,特别是脂肪族硫代葡萄糖苷的含量。从Bn UGT74B1和Bn UGT74C1基因表达中也可以看出,像脱硫-硫代葡萄糖苷糖基化这样的其他硫代葡萄糖苷的生物合成阶段也因为ALA的作用而加强。高质量浓度的ALA处理,通过造成细胞成分损害的光氧化胁迫而对硫代谢和硫代葡萄糖苷合成产生负调控作用。而中质量浓度的ALA促进了硫的获取和同化,并促进硫代葡萄糖苷的生物合成。 相似文献
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[目的]研究镉胁迫下萝卜幼苗的硫代葡萄糖苷含量的变化。[方法]分别用不同浓度的氯化镉(CdCl2)溶液处理盛丰、碧玉春和速白1号3种白萝卜幼苗,研究白萝卜幼苗中抗氧化活性及硫代葡萄糖苷随时间的变化趋势。[结果]镉胁迫下白萝卜幼苗的硫代葡萄糖苷表现出"低浓度促进高浓度抑制"的特点。10和1μmol/L镉胁迫明显促进硫代葡萄糖苷的产生,且10μmol/L促进效果最好。[结论]该研究为了解重金属对白萝卜的作用机制及生化解毒机理奠定了基础,为重金属污染的土壤植物修复提供了理论依据。 相似文献
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《江西农业学报》2022,(3)
以11份菜薹(Brassica campestris L.ssp.chinensis var.utilis Tsen et Lee)品种为材料,采用HPLC法研究了不同基因型菜薹叶片和薹中硫代葡萄糖苷(硫苷)的组分及含量。在11个菜薹品种叶片和薹中都检测到7种硫苷组分,其中3种为脂肪族硫苷,分别为2-羟基-3-丁烯基硫代葡萄糖苷(PRO)、3-丁烯基硫代葡萄糖苷(NAP)和4-戊烯基硫代葡萄糖苷(GBN);4种为吲哚族硫苷,分别为4-羟基-3-吲哚甲基硫代葡萄糖苷(4OH)、3-甲基吲哚基硫代葡萄糖苷(GBC)、4-甲氧基吲哚基-3-甲基硫代葡萄糖苷(4ME)和1-甲氧基-3-吲哚基甲基硫代葡萄糖苷(NEO)。不同基因型菜薹间硫苷总含量和单一硫苷含量均差异显著。薹中4号菜薹的硫苷含量最高,达50.09μmol/100 g FW;叶片中2号菜薹的硫苷含量最高,达34.17μmol/100 g FW。对于同一基因型菜薹,薹中的硫苷含量均显著高于叶片中的,脂肪族硫苷含量显著高于吲哚族硫苷的。NAP和PRO是菜薹中含量较高的硫苷组分。 相似文献
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高温短时蒸汽处理对玛咖黑芥子酶活性及品质的影响 总被引：3，自引：0，他引：3
【目的】为了提高玛咖贮藏和加工后芥子油苷的含量及改善品质,探讨高温蒸汽烫漂在玛咖块根黑芥子酶钝化上的应用效果。【方法】通过单因素试验考察蒸汽压力、蒸汽处理时间和载料量3个主要因素对玛咖块根黑芥子酶活性及芥子油苷含量的影响,在单因素试验的基础上,以黑芥子酶活性以及芥子油苷含量为响应值,进行Box-Behnken中心组合试验。【结果】利用响应曲面法分析优化最佳蒸汽处理工艺条件为：蒸汽压力0.35 MPa,蒸汽处理时间50 s,载料量为240 g,在此条件下玛咖块根黑芥子酶相对酶活性降低到1.96%,且保留的芥子油苷含量达到0.98%。在达到烫漂终点的要求下,高温短时蒸汽处理的最佳工艺与热水烫漂、微波烫漂相比,营养成分保留较多。【结论】高温短时蒸汽处理能有效钝化玛咖块根黑芥子酶活性,且处理后感官品质较佳,与常规烫漂工艺相比,优势较明显。 相似文献
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Sulforaphane is a type of sulfur-containing isothiocyanates hydrolyzed from glucosinolates by myrosinase found in Brassica plants. Sulforaphane is a naturally occurring inducer of phase II enzymes in human and animal bodies to detoxify cancer-causing chemicals. Glucoraphanin is the precursor of sulforaphane and its content is greatly influenced by plant species and genotype, plant organs, pre-harvest factors, and post-harvest processing, thus sulforaphane formation is also directly influenced. Here, we review the formation mechanism of sulforaphane and the factors influencing sulforaphane formation. In the end, the future directions are also discussed. 相似文献
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黑芥子酶是一类催化芥子油苷水解的同工酶, pyk10 是一个在拟南芥根和下胚轴中特异表达的黑芥子酶基因。本研究用PCR方法从拟南芥Columbia生态型基因组中克隆了pyk10 基因启动子片段。序列分析表明:该片段全长1 454 bp,与已发表的pyk10 启动子序列(GenBank accession no. AJ292756)同源性达98%。该启动子片段中含有多种其他植物基因启动子中发现的通用启动子元件,并含有器官和组织特异性转录因子的结合位点ACGT-、CANNTG-、GATA-以及I盒等顺式元件。 相似文献
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Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) 
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下载免费PDF全文 PAN Yan XU Yuan-yuan ZHU Xian-wen LIU Zhe GONG Yi-qin XU Liang GONG Mao-yong LIU Li-wang 《农业科学学报》2014,13(9):1877-1888
Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the related promoter and genomic DNA sequences as well as expression profiles of myrosinase gene remain largely unexplored in radish (Raphanus sativus). In this study, the 2 798 bp genomic DNA sequence, designated as RsMyr2, was isolated and analyzed in radish. The RsMyr2 consisting of 12 exons and 11 introns reflected the common gene structure of myrosinases. Using the genomic DNA walking approach, the 5′-flanking region upstream of RsMyr2 with length of 1 711 bp was successfully isolated. PLACE and PlantCARE analyses revealed that this upstream region could be the promoter of RsMyr2, which contained several basic cis-regulatory elements including TATA-box, CAAT-box and regulatory motifs responsive to defense and stresses. Furthermore, recombinant pET-RsMyr2 protein separated by SDS-PAGE was identified as myrosinase with mass spectrometry. Real-time PCR analysis showed differential expression profiles of RsMyr2 in leaf, stem and root at different developmental stages (e.g., higher expression in leaf at cotyledon stage and lower in flesh root at mature stage). Additionally, the RsMyr2 gene exhibited up-regulated expression when treated with abscisic acid (ABA), methyl jasmonate (MeJA) and hydrogen peroxide (H2O2), whereas it was down-regulated by wounding (WO) treatment. The findings indicated that the expression of RsMyr2 gene was differentially regulated by these stress treatments. These results could provide new insight into elucidating the molecular characterization and biological function of myrosinase in radish. 相似文献
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《农业科学学报》2017,(9)
Plant structures and chemicals, which are developed from the shoot apical meristem(SAM), form the main barriers to insect feeding. A plant chimera containing cells of different genetic origins in the SAM will be morphologically and chemically different compared with the parents and thus may result in differential resistance to herbivores. In this study, we explore if particular elements of plant resistance are localized in one of the layers of SAM; the replacement of one cell layer in a chimera may be linked to change of a single resistance trait to herbivores. The morphology and glucosinolate profiles of two periclinal chimeras(labeled as TTC and TCC, respectively) and grafted parents tuber mustard(labeled as TTT) and red cabbage(labeled as CCC) were compared and the performance of whitefly(Bemisia tabaci) in host selection, oviposition preference and development were assessed under controlled conditions. Both chimeras possessed leaf trichomes as parent tuber mustard TTT, however, TTC had significantly more trichomes than TCC and parent TTT. Leaf wax content of both chimeras was intermediate between the two parents. Five aliphatic and two indole glucosinolates were detected in both chimeras, whereas three aliphatic glucosinolates(3-methyl-sulfinylpropyl, 4-methyl-sulfinylbutyl and 2-hydroxy-3-butenyl) were not detected in tuber mustard, and one aliphatic glucosinolate(3-butenyl) was not detected in red cabbage. Unexpectedly for a chimera, the quantities of two aliphatic glucosinolates(3-methyl-sulfinylpropyl and 4-methyl-sulfinylbutyl) in both TTC and TCC were 3-to 5-fold higher than parents. In olfactory preference assays, B. tabaci showed preference to CCC, followed by TCC, TTC and TTT, and number of eggs laid showed the same pattern: CCCTCCTTCTTT. Interestingly, more whiteflies landed on TTT plants than the other three types in a free choice experiment and the developmental duration from egg to adult was the shortest on TTT and increased in the order TTTTTCTCCCCC. Our results indicate plant defenses traits of leaf waxes, trichomes and glucosinolates are not controlled by one cell layer of SAM, but are influenced by interactions amongst cell layers. The overall findings suggest that periclinal chimera systems can be a valuable approach for the studyof plant-insect interactions and may also be useful for future resistance breeding. 相似文献
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芥蓝黑芥子酶基因的克隆与原核表达 总被引：5，自引：1，他引：4
根据GenBank上已公布的植物黑芥子酶基因序列设计引物,采用RT-PCR和3′/5′-RACE相结合的方法,从芥蓝中克隆出黑芥子酶基因全长cDNA序列.序列全长为1832 bp,开放阅读框为1647 bp,编码549个氨基酸,含有糖基水解酶家族Ⅰ(Glyco_hydro_1)活性位点,GenBank登录号为DQ767973.该核苷酸序列与十字花科其他植物的黑芥子酶基因序列具有较高的同源性,同油菜、萝卜、芥菜和大白菜的同源性达90%以上.利用pET28-α(+)构建芥蓝黑芥子酶基因的原核表达载体,转入大肠杆菌BL21,经IPTG诱导表达,SDS-PAGE检测表明表达蛋白的分子质量约为65 ku. 相似文献