银杏叶片可溶性蛋白质测定非直线性原因分析

曾报告用考马斯亮蓝法测定银杏叶片可溶性蛋白质含量缺乏直线性,猜想其可能具有较高的蛋白水解酶活性。研究表明,银杏叶片蛋白水解酶用ｐＨ７．０缓冲液提取时活性最高,酶活最适ｐＨ５．５~６．０,因而不能解释ｐＨ８．３缓冲液提取出的蛋白质含量下降的原因。比较不同的蛋白质测定方法表明,双缩脲法测得的ＯＤ５４０值相当稳定,而考马斯亮蓝法测定的ＯＤ５９５值随时间延长而升高;同样比色５ｍｉｎ,双缩脲法测得的蛋白质浓度比考马斯亮蓝法高２０~３０倍,说明银杏叶片蛋白质不能迅速溶解于酸性考马斯亮蓝显色液是其测定非直线性的主要原因。     

药用真菌粗多糖蛋白含量测定方法

以药用真菌姬松茸(Agaricus blazei)、鸡腿菇(Coprinus comatus)、猴头(Hericium erinaceus)、灰树花(Grifola frondosa)和灵芝(Ganoderma lucidum)的水提多糖为材料,采用二喹林甲酸法、考马斯亮蓝法、福林-酚法、紫外分光光度法、离子色谱法和传统凯氏定氮法对其中的蛋白含量进行测定和比较分析。结果表明,不同测定方法获得的蛋白含量测定值有差异;笔者推荐使用离子色谱法进行精确蛋白含量测定,采用福林-酚法进行快速趋势比较测定。     

不同产地黑木耳水溶性多糖与蛋白质含量测定比较

为了研究不同产地黑木耳中水溶性多糖和蛋白质含量的差异,采用硫酸-苯酚法和考马斯亮蓝法分别测定了东北、云南丘北、金江、临沧、凤庆五个不同产地黑木耳中的可溶性多糖和蛋白质的含量。结果表明:在供试样品中,东北黑木耳水溶性多糖含量最高达到了5.95%;金江木耳蛋白质含量最高为0.625%,二者均有较高的营养价值。     

不同产地香菇中糖和蛋白质的含量比较

比较5种不同产地香菇中多糖和蛋白质含量,评价香菇质量。采用反向滴定法和考马斯亮蓝法分别测定香菇水提液中香菇多糖和蛋白质的含量。结果表明,多糖含量:河北平泉香菇黄山野山珍香菇湖北神农架野生香菇福建宁德香菇吉林长白山香菇;蛋白质含量:吉林长白山香菇黄山野山珍香菇湖北神农架野生香菇福建宁德香菇河北平泉香菇。     

灰树花多糖提取工艺优化及多糖冲剂的研制

采用正交试验法优化了微波辅助提取灰树花多糖的条件，研究了多糖冲剂制备工艺，并用苯酚-硫酸法对灰树花多糖冲剂中多糖含量进行了测定。结果表明，以水为溶剂，灰树花多糖最佳提取条件为：提取时间10min，微波功率为80％（全功率为800W），液料比为25：1。灰树花多糖冲剂配方为：灰树花粗多糖：可溶性淀粉：甜蜜素为1：4：0．04，冲剂中多糖含量为6．988％．     

双孢蘑菇子实体多糖的提取及其对癌细胞的抑制

双孢蘑菇子实体（Ａｇａｒｉｃｕｓｂｉｓｐｏｒｕｓ）经热水提取，乙醇沉淀，胰蛋白酶消化，Ｓｅｖａｇ法去除蛋白质等处理，得多糖。经紫外可见光分光光度计测定无紫外吸收峰，为较纯多糖。并研究了其对人肝癌ＳＭＭＣ－７７２１细胞生长曲线、有丝分裂指数及线粒体活性的影响，结果表明该多糖对体外培养的人肝癌ＳＭＭＣ－７７２１细胞增殖有一定抑制作用，抑癌实验证明该多糖对小鼠Ｓ－１８０实体瘤亦有一定抑制作用。     

灰树花子实体多糖中重金属含量测定和去除方法的研究

灰树花真菌在栽培过程中往往蓄积重金属,使其含量超标。试验通过对2种重金属去除方法的考察,建立了去除灰树花子实体多糖中重金属的方法。方法：分别采用EDTA和阳离子交换树脂法去除灰树花子实体多糖中的重金属,根据GB／5009m的规定,采用原子吸收分光光度法和原子荧光光度法对去除前后灰树花粗提物中铅,镉,铜,汞,砷等重金属的含量进行测定。结果：732型阳离子树脂法对灰树花子实体多糖的脱重金属效果优于EDTA法。结论：732型阳离子树脂法可用于灰树花子实体多糖中重金属的去除,方法简便,产品收率高且价格便宜。     

蒽酮-比色法测定灰树花多糖条件的研究

为了探讨蒽酮-比色法测定灰树花多糖含量的最佳条件，试验采用沸水水浴不同时间、冰水冷却后不同静置时间的前处理方法测定灰树花提取物的总糖含量。采用酒精浓度为65％、70％、75％、80％、85％、90％、95％、100％，酒精倍数为10倍、25倍、40倍、50倍的稀释浓度，以蒽酮-比色法测定灰树花提取物还原糖含量。结果：沸水水浴不同时间处理对样品总糖含量的影响较大，沸水浴6min后，冷却所测定的总糖含量最高，总糖含量为73．74％。静置时间的长短对吸光值影响不大，在静置4h内测定值稳定性好；不同酒精倍数及不同酒精浓度对样品的还原糖测定结果影响较大。加入50倍酒精、浓度为80％的处理，所测得的还原糖含量最高为15．21％。因此，在灰树花提取物还原糖含量测定时，采用80％的酒精定容至50倍为宜。     

竹荪多糖的分离纯化和鉴定

人工栽培的竹荪子实体干品经热水提取，乙醇沉淀，蛋白酶法和Ｓｅｖａｇ法相结合去蛋白，ＤＥＡＥ－Ｓｅｐｈａｄｅｘ Ａ－２５和Ｓｅｐｈａｄｅｘ Ｇ－２００柱纯化，得到竹荪多糖，命名为ＤＩ。用聚丙烯酰胺凝胶电泳和葡聚糖凝胶柱层析，分析表明，该多糖为均一物质，竹荪多糖的分子量为１４４０００。经紫外扫描，无核酸和蛋白质的特征吸收峰，红外扫描表明有典型的多糖吸收峰。该多糖经纸层析和气相色谱分析表明：其单糖组成为     

酸柚硼毒害蛋白质双向电泳技术体系的优化

以硼毒害下酸柚实生苗的根和叶为试验材料,对双向电泳技术中的样品制备方法、蛋白上样量、凝胶染色方法等条件进行了优化,建立了适用于柑橘蛋白质组研究的双向电泳技术体系。结果表明,使用改良后的酚抽法提取柑橘总蛋白效果较好,所获蛋白在2-DE图谱上获得的点数较多;当采用pH 4～7 IPG胶条、上样量为1.5 mg时,蛋白点分布广,点数最多;使用胶体考马斯亮蓝染色法,可得到背景清晰、重复性好、蛋白点分辨率较高的双向电泳图谱。该体系为开展柑橘硼胁迫的蛋白质组学研究提供了技术基础。     

灰树花深层发酵及菌丝体成分分析研究

本文报道了灰树花菌丝体的深层发酵及有效成分提取和分析。分别从菌丝体及发酵清液中提取得到了灰树花多糖和胞外多糖 ,同时还测定了菌丝体成分 :灰树花多糖 7 82 % ,蛋白质 2 1 7% ,总糖 5 7 2 % ,灰分6 0 5 % ,其中菌丝体灰树花多糖及总糖含量明显多于子实体。     

番茄子叶总蛋白双向电泳体系的建立

通过对IPG胶条梯度、上样方式、上样量、聚焦条件等4方面条件的优化,建立了番茄子叶总蛋白双向电泳体系,即采用TCA丙酮沉淀法提取番茄子叶总蛋白,选用24 cm pH3-7NL的IPG胶条,采用水化上样,上样量300 μg,按聚焦程序Ⅰ或Ⅱ聚焦后进行双向电泳分离和银染染色。若采用制备胶,以获取高含量蛋白点,则将上样量提高至1.5 mg,按聚焦程序Ⅲ进行聚焦后进行双向分离,并采用胶体考马斯亮蓝染色。采用该优化的体系可以获得分辨率高、重复性好的双向电泳图谱。     

Characterisation of proteins in the storage organs of Curcuma alismatifolia Gagnep.

Summary

Curcuma alismatifolia rhizomes are exported to world markets for ornamental use. Rhizome size and the number of storage roots affected shoot sprouting, plant growth, and inflorescence yield and quality, indicating an important role for storage food reserves such as the carbohydrates and proteins in these organs. An understanding of the proteins used for growth and development in rhizomes and in storage roots is lacking, so changes in the storage proteins in these organs were investigated at the planting stage, the sprouting stage, the second-leaf stage, the flowering stage, and the dormancy stage. The protein concentrations in both these organs decreased rapidly until the flowering stage in order to support the initial growth of roots and shoots. The intensity of Coomassie Brilliant Blue staining of tissues was stronger in rhizomes than in storage roots. Protein staining was present in the cytosol and in the cell walls in rhizomes, and was intense in small particle-like protein bodies in the cytosol. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles showed that peptides of 10.6 kDa and 12.0 kDa were the two major soluble proteins in rhizomes and were different from the proteins in storage roots.The 10.6 kDa and 12.0 kDa bands contained five peptides and one peptide, respectively, when separated by 2D-PAGE. The amino acid sequences of these six peptides were not homologous to any known proteins using BLASTP or FASTA analysis. The N-terminal amino acid sequences of these two polypeptides did not start with methionine, indicating that they had undergone posttranslational modification. Knowledge of the major storage proteins in the rhizomes and storage roots of C. alismatifolia may provide information to improve N-fertilisation.     

食用菌复合多糖的抗氧化活性研究

摘要：通过采用化学法，分别测定了猴头菇多糖、香菇多糖、银耳多糖、鸡腿菇多糖、灰树花多糖及香菇复合多糖、灰树花复合多糖体外抗氧化活性。结果表明，0．5mg·mL-1的猴头菇多糖、香菇多糖、银耳多糖、鸡腿菇多糖、灰树花多糖与复合多糖能够不同程度地清除DPPH·自由基、羟自由基（·OH）和超氧阴离子自由基．（02-·）能力，具有较好的抗氧化活性，而香菇复合多糖和灰树花复合多糖的总体抗氧化活性优于其组成单糖，其中以灰树花复合多糖对自由基的清除能力最强，其清除率分别为（89．5±1-3）％、（62．4±0．6）％、（47．9±1．0）％。     

两株西藏白肉灵芝菌株特性初探

对西藏林芝地区的白肉灵芝(Ganoderma leucocontextum)菌株G-122和G-241进行鉴别分析,并对其栽培性状、有效成分含量和提取物功能等方面进行研究。基于形态学特性和ITS分析,2个菌株均鉴定为白肉灵芝。二者菌丝对峙培养产生明显拮抗线,菌丝体生物量差异显著(G-241高于G-122),子实体颜色不同(G-241和G-122分别为红褐色和黄褐色),G-241子实体单菇重高于G-122;有效成分含量差异也较大,G-122粗多糖含量高而G-241总三萜含量高;菌株G-122、G-241醇提物对乳腺癌MT-1细胞IC_(50)值分别为265.2和225.6μg/mL。实验结果表明菌株G-122和G-241为不同菌株。     

Ischemia reperfusion-induced proteomic changes in rat skeletal muscle

AIM: To investigate ischemia reperfusion (I/R)-induced proteomic changes in rat skeletal muscle. METHODS: Healthy male Wistar rats were randomly divided into two groups as follows (n=6): sham group and I/R group. I/R of right hind limb was induced by 4 h ischemia followed by 24 h reperfusion. The 2-DE was applied to separate the proteins extracted from skeletal muscle tissue at the end of experiment, followed by Coomassie Brillant blue R-250 staining. Computer image analysis was used to determine the differential expression of proteins between the two groups, and 7 protein spots expressed differentially were picked out and subjected to in-gel digest and MALDI-TOP for identification. RESULTS: 354±13 proteins were detected and the match rate was (78.7±1.4)%. 10 proteins displayed significant changes after I/R, of which, 6 proteins increased and 3 proteins decreased in expression. Moreover, 2 spots in I/R group were observed, only 1 spots of which in control. 5 proteins were identified after mass spectrometry. Mitochondrial aldehyde dehydrogenase (ALDH) precursor, heat shock 27 kD protein (HSP27), an unnamed protein product (increased in I/R group), α-actin (decreased in I/R group), and nuclear transport factor 2 (NTF-2) W7a mutant were found in I/R group. CONCLUSION: I/R injury induced differential proteomic changes in rat skeletal muscle. ALDH, α-actin and HSP27 expression, and NTF-2 mutation are involved in I/R injury.     

Effect of two UV-absorbing greenhouse-covering films on growth and yield of an eggplant soilless crop

The use of UV absorbing films as greenhouse cover material is spreading out in protected cultivation. Although their effects on pest and disease management have received much attention, few studies focus on their effects on the crop. This study aims at assessing the consequences of UV absorbing film on the behaviour and production of an eggplant crop by comparing two different UV absorbing films (0 and 3% UV transmittance) to a standard polyethylene film (5% UV transmittance). Results show that the eggplants grown in the greenhouse with 0% transmission to UV light are about 21% taller and have about 17% higher leaf product (leaf length × width) than the plants grown in the greenhouse with 5% transmission to UV light. Finally, given that the production was slightly increased in quantity (20%) and quality (bigger fruits) in the greenhouse with absence of UV light compared to that with 5% transmission coefficient, it can be concluded that growing soilless eggplant under UV-absorbing material can be achieved with the same or better results as under standard covering material. Any other enhancement that the UV-absorbing film will bring (lower pest and disease impact on the crop, lower pesticide load and costs) will therefore be to the benefit of the grower.     

Botulinum neurotoxin type A heavy chain intervenes in expression of general proteins in spinal cord after unilateral spinal cord injury in rats

AIM: To observe the effect of botulinum neurotoxin type A heavy chain (BoNT/A HC) on the pattern of spinal protein expression by intrathecal injection after spinal cord injury in rats, and to explore the role of BoNT/A HC intervention in spinal protein expression and some of its mechanisms in nerve regeneration after injury. METHODS: The model of unilateral lumbar spinal cord injury was established. The effects of BoNT/A HC intervention at different doses (2 μg, 4 μg, 6 μg and 8 μg) on the general pattern of protein expression in the spinal cord tissues at the injury site and the cranial part adjacent to the injury site was measured and evaluated by SDS-PAGE and Coomassie brilliant blue staining first, and then by two-dimensional SDS-PAGE. RESULTS: The histological structure of the ipsilateral side of lumbar spinal cord showed obvious destruction and degradation, mainly affecting both gray and white matter of the left side of the cord. The result of SDS-PAGE with Coomassie brilliant blue staining from injured spinal cord tissue displayed that the expression of some proteins after one-time BoNT/A HC treatment appeared obviously different from that without BoNT/A HC treatment. Moreover, the pattern of the protein expression affected by BoNT/A HC was similar to that of the normal spinal cord. The more detail information from two-dimensional SDS-PAGE indicated that more than 10 proteins with different molecular weight and isoelectronic points were differentially expressed at day 2 and day 20 after local injection of 6 μg BoNT/A HC. This altered expression actually appeared a tendency toward the pattern shown in normal group. CONCLUSION: The immediate application of BoNT/A HC at the injury site after unilateral lumbar spinal cord injury is able to affect the pattern of local protein expression. The altered protein expression by injury could be reversed back to normal or approximately normal by local BoNT/A HC administration.