茄子果实采后软化过程中细胞壁组分及其降解酶活性的变化

研究了细胞壁组分及其降解酶活性的变化与茄子果实采后软化的关系。结果表明,采后茄子果肉硬度随贮藏时间的延长而不断下降。贮藏期间果肉水溶性果胶(WSP)含量在贮藏前12天不断增加,之后快速下降,而共价结合型果胶(CSP)、半纤维素和纤维素等细胞壁组分含量持续减少。果肉果胶甲酯酶(PME)、多聚半乳糖醛酸酶(PG)和纤维素酶(CX)活性均呈先升高后下降趋势,分别在贮藏至第6、9、12天达到最大值;β-半乳糖苷酶(β-Gal)活性始终保持较高水平,且在整个贮藏期间活性变化不明显。相关性分析结果表明,CSP、半纤维素和纤维素的降解与采后茄子果实软化密切相关,PG和CX在茄子果实采后软化过程中起着重要的作用。     

冷藏对蓝莓果实细胞壁组分及其降解酶活性的影响

以"蓝丰"蓝莓果实为试材,研究了冷藏对蓝莓果实细胞壁组分及其降解酶活性的影响。结果表明:蓝莓果实的软化与离子结合型果胶(ISP)、共价结合型果胶(CSP)含量和多聚半乳糖醛酸酶(PG)、纤维素酶(Cx)、β-半乳糖苷酶(β-Gal)活性关系密切,其中,ISP含量和PG活性与果实硬度呈显著负相关,Cx和β-Gal活性与果实硬度呈极显著负相关,CSP含量与果实硬度呈极显著正相关;冷藏期间,随果实硬度的逐渐下降,CSP含量逐渐降低,ISP含量变化幅度小,PG和β-Gal活性呈上升趋势,Cx活性呈现先下降后上升趋势,3种酶均在贮藏的后期活性急剧上升;在冷藏的30 d,果实的细胞壁组分和相关酶活性变化较小,果实硬度下降缓慢;与采后自然后熟的果实相比,冷藏30 d的蓝莓果实常温货架期间Cx活性和β-Gal活性一直处于较低水平,PG活性高峰延晚出现,Cx活性高峰极显著降低,果胶甲酯酶(PE)活性未出现高峰。可见,冷藏30 d的蓝莓果实细胞壁组分含量及相关酶活性的变化一定程度受到抑制,果实状态保持良好。     

一氧化氮对常温贮藏下芒果果实软化和细胞壁代谢的影响

为了探明一氧化氮(NO)抑制采后芒果软化的作用机理,将"台农"芒果果实在0.25 mmol/L硝普钠(SNP,NO供体)溶液浸泡处理20 min,常温(20±2) ℃贮藏20 d,定期测定果实硬度、细胞壁组分含量、细胞壁水解酶活性。结果表明,与未处理果实相比,SNP处理显著降低贮藏20 d内果实中多聚半乳糖醛酸酶(PG)活性(p<0.05),显著抑制贮藏10 d内果实纤维素酶(CX)(p<0.05)活性,极显著抑制β-半乳糖苷酶(β-Gal)和α-L-阿拉伯呋喃糖苷酶(α-L-Af)活性(p<0.01),但使贮藏15~20 d期间果实CX和β-Gal活性及贮藏第20 d的α-L-Af活性均显著增加(p<0.05)。SNP处理显著抑制贮藏5 d内果胶甲酯酶(PME)活性(p<0.05),但在贮藏10~20 d期间保持较高的PME活性(p<0.05)。此外,SNP处理极显著延缓原果胶和纤维素的降解(p<0.01),减少可溶性果胶含量的增加(p<0.05),从而降低贮藏期间果实硬度的损失。硬度与原果胶、纤维素含量均呈极显著正相关(p<0.01),而与CX活性呈显著负相关(p<0.05),与可溶性果胶含量、PG、β-Gal和α-L-Af活性均呈极显著负相关(p<0.01)。可溶性果胶含量与纤维素含量呈极显著负相关(p<0.01),而与α-L-Af活性均呈显著正相关(p<0.05),与PG和β-Gal活性均呈极显著正相关(p<0.01)。因此,采后SNP处理可以通过调节果实细胞壁降解酶活性,减少细胞壁组分的降解,从而延缓芒果采后软化,延长贮藏期。     

高能电子束辐照对猕猴桃细胞壁降解相关酶活性和基因表达的影响

以‘海沃德’猕猴桃为试材,经剂量0（对照）、300、400和500 Gy高能电子束辐照后,于0~1 ℃、RH 90%~95%冷库中贮藏90 d,研究电子束辐照对果实硬度、细胞壁组分、软化相关酶活性及其基因表达量的影响。结果表明:高能电子束辐照显著维持了果实的硬度,有效抑制了细胞壁骨架物质原果胶和纤维素的分解,延迟了果实后熟软化。同时,辐照抑制了多聚半乳糖醛酸酶（polygalacturonase,PG）、果胶甲酯酶（pectin methylesterase,PME）、β-半乳糖苷酶（β-D-galaetosidase,β-Gal）和纤维素酶（cellulase,Cx）的活性,降低了PG、PME、β-Gal和Cx编码基因的表达。综合认为,以400 Gy高能电子束辐照对抑制细胞壁降解相关酶活性及基因表达,保持细胞结构的完整性,维持贮藏期间果实硬度效果最好。研究结果为高能电子束用于猕猴桃采后保鲜提供理论依据。     

不同成熟度橄榄果实冷藏期间细胞壁代谢对采后冷害的响应特性

研究冷藏期间橄榄果实细胞壁代谢的变化,探讨不同成熟度橄榄果实冷害发生与细胞壁组分含量、细胞 壁降解酶活性的关系。以白露、寒露、立冬、大雪节气时采摘的‘檀香’橄榄果实为材料,在温度（2±1）℃、 相对湿度85%～90%冷库内贮藏,定期测定橄榄果实冷害指数、果肉细胞壁组分含量和细胞壁降解酶活力的变化。 结果表明,不同成熟度橄榄果实冷藏期间冷害发生与其细胞壁组分降解密切相关,冷害指数与离子结合型果胶 （ionic-soluble pectin,ISP）、共价结合型果胶（covalent-soluble pectin,CSP）、半纤维素和纤维素含量呈负相 关;且果胶甲酯酶（pectin methylesterase,PME）、多聚半乳糖醛酸酶（polygalacturonase,PG）、β-半乳糖苷酶 （β-galactosidase,β-Gal）和纤维素酶（cellulase,CEL）等细胞壁降解酶的活力变化不平衡或活力提高是导致冷藏 橄榄果实细胞壁结构解体、细胞壁代谢异常、冷害发生的主要原因。同时,与成熟度Ⅰ、Ⅲ和Ⅶ的橄榄果实相比, 成熟度Ⅴ保持较低的果实冷害指数及冷藏中后期果肉PME、PG、β-Gal和CEL活力,延缓冷藏中后期果肉水溶性果 胶、ISP、CSP、半纤维素和纤维素含量降低。因此认为,成熟度Ⅴ的橄榄果实可较好维持细胞壁结构的完整性, 有效减轻冷害发生。     

苹果发育过程中细胞壁代谢及果肉质地的变化

以质地存在差异的“蜜脆”、“嘎拉”、“倭锦”、“鸡冠”4 个品种的苹果为材料,分析了果实发育过程中硬度、脆度和单果质量的变化,以及细胞壁中果胶、纤维素、半纤维素的含量和多聚半乳糖醛酸酶（polygalacronase,PG）、果胶甲酯酶（pectinmethylesterase,PME）、纤维素酶（carboxymethycellulase,Cx）、β-半乳糖苷酶（β-galactosidase,β-Gal）活性的变化,以期找到品种间质地差异的主要因素。结果表明：果实发育过程中果实硬度降低、脆度变小、感官脆度增大,细胞壁各个组分的含量在4 个品种中均呈下降趋势;在成熟果实中,脆性较好的品种果实中纤维素含量较低,水溶性果胶（water soluble pectin,WSP）含量较高,硬度低的品种果实中离子结合果胶（ionic soluble pectin,ISP）含量较高;PG、β-Gal在高脆品种“蜜脆”、“嘎拉”中活性较高,而Cx、PME活力在各品种间无明显差异。认为WSP、ISP和纤维素含量的差异以及PG和β-Gal活性的差异是形成苹果果实质地差异的关键因素。     

外源草酸对采后‘蜂糖李’果实软化和细胞壁降解的影响

为探讨外源草酸处理对采后李果实软化进程的影响,以‘蜂糖李’果实为试材,采用5 mmol/L草酸(oxalic acid, OA)溶液浸泡处理10 min,以蒸馏水浸泡处理10 min为对照(CK),自然晾干后置于室温(25±1)℃条件下贮藏20 d。分析果实硬度、纤维素、原果胶和可溶性果胶含量的变化以及多聚半乳糖醛酸酶(polygalacturonase, PG)、果胶甲酯酶(pectin methylesterase, PME)、纤维素酶(cellulase, Cx)、β-半乳糖苷酶(β-galactosidase, β-Gal)、α-L-阿拉伯呋喃糖苷酶(α-L-arabinofuranosidase, α-L-Af)和木葡聚糖内糖基转移酶(xyloglucan endotransglucosylase, XET)活性。结果表明,外源OA处理能使‘蜂糖李’果实保持较高的硬度,延缓原果胶和纤维素含量的下降以及可溶性果胶含量的增加,抑制果实PG、PME、Cx、β-Gal、α-L-Af和XET活性上升。果实硬度与原果胶、纤维素、可溶性果胶含量、Cx、β-Gal、XET和α-L-Af活性均...     

微环境气调对蓝莓贮藏期软化的调控作用

为探讨微环境气调对蓝莓贮藏期果实软化的影响,采用自发气调(mMAP)、1-甲基环丙烯(1-MCP)、微环境气调(mMAP+1-MCP)处理蓝莓,以未经处理的蓝莓为对照。将处理过的蓝莓置冰温库(-0.5±0.3) ℃贮藏,分别于贮藏0,20,40 d和60 d时测定果实硬度、细胞壁多糖含量、细胞壁降解酶活性和关键降解酶基因表达量。结果表明:与对照组相比,3个处理组均能显著降低果实的软果率(P < 0.05),其中mMAP+1-MCP处理效果最佳。贮藏60 d时果实硬度显著高于其它处理组(P < 0.05),维持较高的纤维素和原果胶含量,半纤维素在贮藏前期高于其它处理,而可溶性果胶含量在贮藏中前期保持较低水平。分析细胞壁降解酶,mMAP+1-MCP处理组贮藏40 d时纤维素酶(Cx)、多聚半乳糖醛酸酶(PG)和果胶甲酯酶(PME)活性最低,贮藏60 d时β-半乳糖苷酶(β-Gal)和α-L-阿拉伯呋喃糖苷酶(α-Af)活性显著低于其它处理组(P < 0.05)。正交偏最小二乘判别分析(OPLS-DA)表明,mMAP+1-MCP与其它处理组差异性指标为Cx和β-Gal活性。对这两种细胞壁降解酶基因的表达分析结果:mMAP+1-MCP处理可有效延缓蓝莓贮藏过程中Cx和β-Gal基因表达量峰值的出现时间。结论:mMAP+1-MCP形成的微环境气调环境通过延缓Cx和β-Gal基因表达量出峰时间,抑制贮藏后期Cx和β-Gal活性,保持果实的纤维素和原果胶含量,进而减缓果实的软化。     

钙调素拮抗剂三氟啦嗪对辐照蓝莓果实硬度品质特性的影响

为了探讨γ辐照对冷藏蓝莓果实软化的抑制机理,本研究采用2.5 kGy剂量辐照处理“蓝丰”蓝莓,结合钙调素拮抗剂TFP,测定各组蓝莓果实硬度和原果胶、不同溶解性果胶含量、纤维素和半纤维素含量,以及相关水解酶活性在冷藏期间的变化差异,结果显示：辐照能够介导Ca2+信号转导抑制PG和PME活性,降低原果胶和CSP降解为WSP的可能,进而对冷藏蓝莓果实硬度产生积极影响;TFP能够在一定程度上阻断胞内Ca2+信号的转导作用;但是Cx、β-Gal和α-Af对辐照诱导的Ca2+信号响应不积极,辐照后冷藏蓝莓果实4KSF、24KSF和纤维素含量的变化与Ca2+信号转导无关。因此,γ辐照诱发的Ca2+信号对果胶参与的冷藏蓝莓果实软化具有重要的调控作用。     

海藻酸钠/纳米TiO2复合涂膜对采后番木瓜果实软化及细胞壁降解影响

为探究海藻酸钠/纳米TiO₂(sodium alginate/nano-TiO₂,SA/TiO₂)复合涂膜处理抑制采后番木瓜果实软化的作用机理,以“日升10号”番木瓜为试材,采用SA/TiO₂复合涂膜处理,研究采后番木瓜果实硬度、细胞壁组分含量和细胞壁水解酶活性的变化规律。结果表明,SA/TiO₂复合涂膜处理能有效地抑制采后番木瓜果实中多聚半乳糖醛酸酶(polygalacturonase,PG)、果胶甲酯酶(pectin methylesterase,PME)、纤维素酶(cellulase,Cx)和β-半乳糖苷酶(β-D-galaetosidase,β-Gal)活性,减缓原果胶和纤维素的水解,减少可溶性果胶的增加,从而保持较完整的细胞壁结构,维持果实硬度。因此认为,SA/TiO₂复合涂膜处理可延缓采后番木瓜果实的软化进程,提高果实耐贮性。     

酵母细胞破壁技术研究与应用进展

随着啤酒行业的发展,啤酒生产中带来的废酵母数量与日俱增,造成了环境污染,废酵母的再利用成为人们关注的焦点。酵母细胞内具有丰富的营养物质,充分利用这些营养物质需要对酵母细胞进行破壁,因而破壁技术就显得尤为重要。系统论述了当前破碎酵母细胞壁的方法和原理,以及各种方法的优缺点和应用现状,对酵母破壁技术的应用前景进行展望。     

Similar and different domains of chitin synthases 1 and 2 of S. cerevisiae: two isozymes with distinct functions

Saccharomyces cerevisiae produces two chitin synthases (Chs1 and Chs2) encoded by separate genes. Although these enzymes catalyze the same reaction, Chs2 is essential for septum formation whereas Chs1 has a repair function. To determine if these physiological differences are reflected in the enzyme structures, the CHS2 gene was sequenced and compared to that of CHS1. The predicted amino acid sequence of Chs2 shares substantial similarity with that of Chs1 in the carboxyl two-thirds of the protein. The amino one-third segments differ in predicted isoelectric point by almost 5 pH units. It is suggested that the similar regions are related to common catalytic function. The unrelated regions may be involved in regulation or localization of the respective enzymes. CHS1 and CHS2 are unlinked but may have arisen from the duplication of an ancestral gene.     

Identification of Candida albicans wall mannoproteins covalently linked by disulphide and/or alkali‐sensitive bridges

This paper describes the results obtained by analysing the human pathogen Candida albicans cell wall subproteome by mass spectrometry, using extraction procedures aimed at releasing proteins bound by disulphide bridges (RAE‐CWP) or alkali‐labile ester linkages (ALS‐CWP). Ten of the total proteins released from the wall by β‐ME and/or NaOH contained a potential signal peptide, lacked a GPI cell wall hydrophobic C‐terminal domain and were identified as true wall proteins by in silico analysis, whereas four additional proteins were identified as bound to the plasma membrane. The results surprisingly demonstrated that, in addition to the expected RAE‐CWP and ALS‐CWP proteins, 16 GPI proteins were bound to the wall by disulphide or alkali‐sensitive bonds, since they were released by β‐ME and/or NaOH. The biological significance of these results is discussed in relation to the added complexity of the organization of the proteins in the C. albicans cell wall. Copyright © 2014 John Wiley & Sons, Ltd.     

Binding of heterocyclic amines by yeast cells and their fractions

The binding of mutagenic pyrolysis products to cells of 50 yeast strains and their cell fractions was investigated. Cells of all yeast strains effectively bound 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). Cell walls (CW), and cell wall glucan and mannan (5 mg in each case) showed the highest binding of Trp-P-1 (50 μg ml^?1); glucan adsorbed virtually all of the Trp-P-1. Cytoplasm also showed some binding but was much less effective. Glucans also bound well with 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-3,8-dimethylimidazo 4,5-quinoxaline (MeIQX) much more than CW, but 2-amino-5-phenylpyridine (Phe-P-1) and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MelQ) were not effectively bound. The quantity of IQ, MeIQ, Phe-P-1 and MeIQX bound was dependent on the strain of yeast. The mutagenic pyrolysis products bound to cells were effectively extracted by aqueous methanol, ammonia (50 g litre^?1) and alcohol, but not by water. The binding was pH dependent and inhibited by metal salts. When yeast cells were heated to 100° for 15 min, the binding of Trp-P-1 decreased by about 30% but Saccharomyces cerevisiae 50 heated to 100° did not differ much from untreated cells in its binding ability.     

细胞壁组分及酶活性与清见果实囊衣绵韧的相关性

以清见杂柑为研究材料,进行定点和定期的跟踪测定,探究清见果实囊衣中果胶类物质、纤维素、半纤维素、木质素含量和果胶甲酯酶、多聚半乳糖醛酸酶、纤维素酶活性在花后120～300 d的动态变化情况,及其与清见果实囊衣绵韧程度的相关性。结果表明：囊衣胞壁物质含量在成熟后期均有下降,多聚半乳糖醛酸酶和纤维素酶活性在后期处于上升趋势,果胶甲酯酶则相反;较高水平的水溶性果胶和低水平的木质素、纤维素、半纤维素有利于清见果实良好化渣性的形成;从花后210～300 d,细胞壁中多聚半乳糖醛酸酶、纤维素酶活性高低是决定囊衣质地绵韧程度的关键因子。     

预处理对增加红富士苹果细胞壁物质降解和出汁率的影响

为了通过简便易行的预处理增强果实细胞壁物质降解,提高苹果果实出汁率。文中测定了500mg/L乙烯利(E_1)和100mg/L乙烯利配合60℃热水(E_2)处理,对红富士果实堆放期细胞壁酶活性、细胞壁物质含量和出汁率的影响。结果表明:E_1使其果胶甲酯酶(PME)、纤维素酶(CS)活性高峰分别比对照提前6、3d,多聚半乳糖醛酸酶(PG)活性在前15d持续高于对照,木聚糖酶(Xyl)活性基本不变;处理12d后,果实细胞壁多糖中果胶类多糖、半纤维素类多糖和细胞壁残渣多糖含量分别下降到对照的80.1%、70.4%和75.7%;处理后9～18d,红富士苹果的出汁率较对照提高2.3%～4.0%;E_2抑制了果实PME活性,却普遍增强了0～6d内PG、Xyl、CS活性,亦促进了各类细胞壁物质的降解,0～9d内果实出汁率较对照高3.2%～7.1%。     

Ripening‐related changes in the cell walls of olive (Olea europaea L.) pulp of two consecutive harvests

Olive fruits, harvested in two consecutive seasons at green, cherry and black stages, were used to study compositional changes in the cell walls during ripening. Ripening‐related changes in both harvests were characterised mainly by an increase in the solubilisation of pectic and hemicellulosic polysaccharides, an increase in the relative amount of arabinose in pectic polysaccharides and a decrease in the degree of methylesterification of pectic polysaccharides. Further to degrading processes, the data obtained suggest the synthesis of new polysaccharides. The analysis of olive cell wall phenolics showed mainly the presence of p‐coumaric acid, which increased in one harvest, whereas in the other the values did not differ. The samples of the second harvest, although presenting green, cherry and black colours, had less distinct ripening characteristics than those of the previous harvest. Different activity levels of polyphenol oxidase, polygalacturonase and pectin methylesterase might have contributed to the differences observed between the two harvests. The results showed the distinct extension of ripening‐related changes in the cell walls of the two harvests, indicating that the olive colour, although characteristic of the stage of ripening, cannot be strictly used for its evaluation and definition. Copyright © 2006 Society of Chemical Industry     

草莓果实成熟过程中细胞壁组分变化的研究

目的研究草莓果实发育成熟过程中细胞壁成分的变化。方法以丰香和红丰草莓为试材，选择小而绿、大而绿、白熟、红熟、全熟5个时期采收，测定细胞组分含量。结果与结论随果实发育成熟，细胞壁中可溶性果胶和半纤维素增加，而离子结合果胶和共价结合果胶及纤维素减少，导致草莓果实成熟软化。     

Cell surface hydrophobicity and colony morphology of Trichosporon asahii clinical isolates

Trichosporon asahii is a pathogenic basidiomycetous yeast. Individual strains of T. asahii have different colony morphologies. However, it is not clear whether cell surface phenotypes differ among the colony morphologies. Here we characterized the cell surface hydrophobicity and analysed the carbohydrate contents of the cell surface polysaccharides in T. asahii clinical isolates with various colony morphologies. Among the three distinctive colony morphologies obtained from one clinical isolate, the white‐type morphology exhibited higher hydrophobicity. The hydrophobicity of heat‐killed T. asahii cells was greatly reduced after periodate oxidation of the cell surface carbohydrates. Furthermore, the cell wall and extracellular polysaccharide components differed among the morphologies. Our results suggest that T. asahii cell surface hydrophobicity is affected by cell surface carbohydrate composition. Copyright © 2016 John Wiley & Sons, Ltd.