毛花猕猴桃中的两个新三萜化合物

从毛花猕猴桃(Actinidia eriantha Benth.)的根中,分离到β-谷甾醇,胡萝卜甙,熊果酸,2α,3α,24-三羟基-12-烯-28-乌苏酸,同时还得到两个新的三萜化合物,命名为毛花猕猴桃酸A(eriatic acid A)和毛花猕猴桃酸B(eriantic acid B)。根据光谱数据及化学反应证明,其结构分别为24-乙酰氧基-2α,3α-二羟基-12-烯-28-乌苏酸和2β,3β,23-三羟基-12-烯-28-乌苏酸。     

毛花猕猴桃两新变种

<正> 秃果毛花猕猴桃 新变种 (图版1) Actinidia eriantha Benth. var. calvescens C. F. Liang, var. nov. A typo differt baccis cylindricis, majoribus, circa 5 cm. longis, tomentosis non velutinis. Guangxi: Guilin, in the Research Orchard of Guangxi Institute of Botany, May 9, 1987, C. F. Liang 34511 (Type, IBK).     

毛花猕猴桃根中的一个新三萜化合物

从毛花猕猴桃(Actindia eriantha Benth)的根中分离到三个结晶成分,根据光谱数据(MS,IR,~(12)CNMR,~1HNMR)和化学证明,R_6的结构为2α,3β,24-三羟基-12-烯-28-乌苏酸是一新化合物,其余两个成分分别为二十四碳酸和葡萄糖。     

毛花猕猴桃原生质体再生植株

从毛花猕猴桃（Ａｃｔｉｎｉｄｉａ ｅｒｉａｎｔｈａ Ｂｅｎｔｈ．）试管培养的实生苗新展开叶片分离的原生质体,培养在液体ＭＳ（除去ＮＨ４ＮＯ３）附加２,４－Ｄ １．０ ｍ ｇ／Ｌ和葡萄糖０．４ ｍ ｏｌ／Ｌ的培养基上。培养３周后植板率达到１９．４％ 。在未添加新鲜培养基的情况下,原生质体再生的细胞可持续分裂,并于３个月时长成２ ｍ ｍ 大小的愈伤组织。将该愈伤组织转移到附加玉米素０．５ ｍ ｇ／Ｌ和ＩＡＡ ０．１ ｍ ｇ／Ｌ的固体ＭＳ培养基上,分化出苗。试管苗经诱导生根,长成完整小植株     

毛花猕猴桃一新变型

<正> 白色毛花猕猴桃 新变型 Actinidia eriantha Benth. form. alba C. F. Gan, f. nov. A typo receditfloribus albis. Zhejiang: Qingyuan-Xian, May 22, 1982, G. F. Gan 101 (Type, con-     

中华猕猴桃和毛花猕猴桃果实碳水化合物及维生素Ｃ的动态变化研究

分别对中华猕猴桃（Actinidia chinensis）黄肉品种‘金桃’和毛花猕猴桃（Actinidia eriantha）品系‘6113’果实生长发育过程中碳水化合物及维生素C的动态变化进行了系统研究。结果表明,中华猕猴桃‘金桃’和毛花猕猴桃‘6113’果实的可溶性固形物（SSC）含量均于谢花后146d内保持相对平稳,而后开始上升;此时,两物种果实的淀粉含量均上升到最大值,之后两者均开始下降。两者糖含量的变化与SSC相似,且中华猕猴桃‘金桃’果实糖含量进入快速增长期的时间比毛花猕猴桃‘6113’早1个月。两者果实Vc含量的变化趋势相似,均于7月上中旬达到一个高峰,以后随着果实的生长发育,含量下降,‘金桃’于8月14日降至最低值,‘6113’于9月13日降至最低值;两者的Vc含量降到最低值后均缓慢上升,到果实完全成熟期（树上自然软熟期）回升到第二个峰值。‘6113’果实的Vc含量在完全成熟期的峰值远远高于7月上旬的高峰值。对‘金桃’和‘6113’果实碳水化合物及Vc含量方差分析表明,两者的可溶性固形物、淀粉和总糖没有明显差异,而毛花猕猴桃‘6113’的Vc含量显著高于中华猕猴桃‘金桃’。     

毛花猕猴桃根中的一个新三萜

毛花猕猴桃(Actindia eriantha Benth)系猕猴桃属落叶藤本植物。我国民间用其根治疗胃癌、鼻咽癌、乳癌等多种疾病。有关毛花猕猴桃根的化学成分研究尚未见报道。我们从广西龙胜县城附近采集到毛花猕猴桃根,首次进行了化学成分的研究,已从中分离得到多种单体成分,经光谱测定和化学反应确定了其中的五个结构,它们是β-谷甾醇,熊果酸,胡萝卜甙,2α,3α,24-三-羟基-12-烯-28-乌索酸(AE5)和一个新的三萜酸:毛花猕猴桃酸A。其它单体的结构将另文发表。     

短小芽孢杆菌碱性蛋白酶的提纯和性质的研究

由短小芽孢杆菌(Bacillus pumilus)209产生的胞外碱性蛋白酶的粗酶制剂(5×10 4u／g),经硼砂NaOH缓冲液抽提,硫酸铵沉淀,Sephadex G一25脱盐,DEAE-纤维素柱层析,冷冻干燥获得部分纯化的酶。纯酶的活力为199x10u/g,比活力提高2,6倍。此酶的最适pH8．5—9．0,在pH6一10之间稳定,因此是属于微碱性蛋白酶“’。最适温度为50℃,在50℃以下稳定,在60℃处理10分钟活力损失95％。0.003村c丑件的存在对酶的热稳定性和pH稳定性均有显著提高。Ag+,Cu2+,Fe2+,Hg+,Zn2+ 对酶有抑制作用;Mn2+有明显的激活作用;1×10-3M的PCMB,IAA,O-PTH,PMSF对活力无影响;1×10-3M EDTA对酶有30％的抑制作用;在40℃用NBS(1×10-4M)、DFP(2．5mM)对醢进行10分钟处理,酶活力完全损失。此酶能水解多种天然蛋白,如酪蛋白、血红蛋白、蛇毒蛋白等,不水解鱼精蛋白和溶菌酶。以酪蛋白为底物测得的km值为0．66×10-2g／ml。在pH7．3进行圆盘凝胶电泳,虽然呈现九条带,但均具有大小不等的蛋白酶活力。     

地衣芽孢杆菌碱性蛋白酶的研究Ⅱ碱性蛋白酶的提纯和性质研究

地衣芽孢杆菌（Ｂａｃｉｌｌｕｓ ｌｉｃｈｅｎｉｆｏｒｍｉｓ）Ｂ．Ｌ ＪＦ－１ｄ三级发酵的发酵液经离心去菌体,（ＮＨ４）２ＳＯ４分段盐析,透析后进行Ｓｅｐｈａｄｅｘ Ｇ－１００柱层析得粗酶制剂。比活力从１８７８Ｕ／ｍｇ提高到６７９５Ｕ／ｍｇ,酶活力回收率为３５．３％。该酶水解酷蛋白的最适反应温度为５５℃,最适ｐＨ为１０．５,具有较高的热稳定性,对ＳＤＳ有较强的耐受性。     

Plant regeneration from in vitro-cultured seedling leaf protoplasts of Actinidia eriantha Benth

Newly expanded in vitro leaves of Actinidia eriantha were used for protoplast isolation. Protoplasts were cultured in liquid MS medium (lacking NH₄NO₃) supplemented with 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.4 M glucose. The plating efficiency after 3 weeks of culture was 19.4%, and calli were recovered without addition of fresh medium. These calli regenerated shoots on transfer to MS medium containing 2.28 μM zeatin and 0.57 μM IAA (indole-3-acetic acid). Regenerated shoots were rooted by immersion in 20 ppm IBA (indole-3-butyric acid) solution before culturing on half-strength MS medium lacking growth regulators. Somaclonal variation, in terms of chromosome number and nuclei per cell of protoplast-derived plants, was estimated. Received: 15 March 1997 / Revision received: 27 January 1998 / Accepted: 7 March 1998     

彩叶草红色素的理论性质

天然色素可从动植物相应组织中提取。从吊竹梅（Zebrina pendula Schnizl．）中已得到非常稳定的天然色素,从Acalypha uilkesiana中得到大量的花青苷色素,从Setcrease purpurea中也得到非常稳定的天然紫红色素。彩叶草（Coleus blumei Benth）含有大量类黄酮物质,目前,对其色素的理化性质没有详细的报道。本文探讨彩叶草红色素的理化性质,旨在为该色素的开发应用提供科学依据。     

Purification and properties of a protease inhibitor from crayfish hemolymph

A protease inhibitor from the hemolymph of crayfish, Astacus astacus, has been purified by differential centrifugation, acid precipitation and preparative isoelectric focusing. The inhibitor was apparent homogenous in SDS-electrophoresis and had a molecular weight of 23,000. pI was determined to be 4.7 by isoelectric focusing. No inhibitory activity was lost when the inhibitor was incubated in a pH range of 1–11.5. The purified inhibitor was heat stable. Urea (6 m) had no effect upon the inhibitor. The inhibitor was active against subtilisin and a partly purified protease from the fungus Aphanomyces astaci. Pronase was slightly inhibited whereas trypsin, chymotrypsin, papain, Arthrobacter protease, and extracellular proteases from the fungi Aphanomyces stellatus and A. laevis were unaffected. The importance of protease inhibitors in pathogenesis between the parasitic fungus, A. astaci, and its crayfish host, A. astacus is discussed.     

Trypsin-like protease from soybean seeds. Purification and some properties

An enzyme was purified from soybean seeds mainly by repeated ion-exchange chromatography using benzoyl-L-arginine p-nitroanilide (BAPA) as a substrate. The purified enzyme was homogeneous as judged by disc gel electrophoresis. The molecular weight was estimated as 59,000 by gel filtration. The enzyme was most active toward BAPA between pH 8 and 10. The enzyme was inactive toward protein substrates but hydrolyzed synthetic substrates and oligopeptides exclusively at the carboxyl side of L-arginine and L-lysine. Kinetic studies using synthetic substrates showed that, on the basis of Vmax/Km, the enzyme preferentially hydrolyzed amide substrates over ester substrates. Benzoyl-L-arginine 4-methylcoumaryl-7-amide (Bz-Arg-MCA) was the best substrate. The enzyme was strongly inhibited by diisopropylfluorophosphate (DFP), tosyl-L-lysine chloromethyl ketone (Tos-Lys-CH2Cl), leupeptin, and antipain. p-Chloromercuribenzoate (PCMB) was only partially inhibitory. Various protein inhibitors of trypsin such as soybean trypsin inhibitor were ineffective. From the primary specificity and susceptibility to chemicals, the enzyme can be said to be a trypsin-like serine protease. Although the physiological role of the enzyme is unclear, it seems likely that it is involved in limited hydrolysis of certain physiological peptides during processing.     

Transformation of Actinidia eriantha: A potential species for functional genomics studies in Actinidia

Protocols were developed for regeneration and Agrobacterium-mediated transformation of Actinidia eriantha Benth. A. eriantha has a number of features that make it a useful tool for functional genomics in Actinidia: the vines are relatively small and non-vigorous in nature, flowers form all over the vine including on lower axillary branches and the species flowers prolifically in greenhouse conditions. Flowering and fruiting of transgenic A. eriantha plants was obtained within 2 years of transformation in a containment greenhouse. GUS (β-glucuronidase) activity indicating stable expression of the uidA gene was observed in leaf, stem, root, petal and fruit tissues. Molecular evidence for incorporation of transgenes into the A. eriantha genome was obtained by PCR and DNA gel blot analysis. Inheritance of transgenic phenotypes was demonstrated in seedling progeny. Functional genomic studies in kiwifruit have been initiated using transgenic A. eriantha plants. Communicated by F. Sato     

牛耳枫中生物碱的研究(英文)

应用各种色谱方法从牛耳枫的茎叶乙醇浸提物中分离得到了8个生物碱,利用波谱(ESI-MS,1HNMR,13C NMR)技术及理化性质分别鉴定为Methyl homosecodaphniphyllate(1),Daphnezomine M(2),Caldaphni-dine E(3),Calyciphylline F(4),Calyciphylline B(5),Deoxycalyciphylline B(6),Daphnicyclidin H(7)和Macropodumine C(8)。     

Purification and characterization of an alkaline protease from Pseudomonas aeruginosa MN1

An alkaline protease produced by Pseudomonas aeruginosa MN1, isolated from an alkaline tannery waste water, was purified and characterized. The enzyme was purified 25-fold by gel filtration and ion exchange chromatography to a specific activity of 82350 U mg⁻¹. The molecular weight of the enzyme was estimated to be 32000 daltons. The optimum pH and temperature for the proteolytic activity were pH 8.00 and 60°C, respectively. Enzyme activity was inhibited by EDTA suggesting that the preparation contains a metalloprotease. Enzyme activity was strongly inhibited by Zn²⁺, Cu²⁺ and Hg²⁺(5 mM), while Ca²⁺ and Mn²⁺ resulted in partial inhibition. The enzyme is different from other Pseudomonas aeruginosa alkaline proteases in its stability at high temperature; it retained more than 90% and 66% of the initial activity after 15 and 120 min incubation at 60°C. Journal of Industrial Microbiology & Biotechnology (2000) 24, 291–295. Received 09 June 1999/ Accepted in revised form 24 January 2000     

Development of microsatellite markers in an amphicarpic species,Amphicarpaea edgeworthii Benth. (Leguminosae)

Eight polymorphic simple sequence repeats (SSRs) markers were developed for Amphicarpaea edgeworthii Benth., an amphicarpic species in East Asia. The low level of heterozygosity in populations and the high level of population differentiation found in this study suggest that A. edgeworthii has a mixed mating system that is predominantly selfing. The findings also indicate that the new markers can be used in genetic analyses of the mating system and the estimation of the construction of progeny populations contributed by chasmogamously and cleistogamously produced seeds.     

Oxidant and SDS-stable alkaline protease from Bacillus clausii I-52: production and some properties

AIMS: An investigation was carried out on an oxidative and SDS-stable alkaline protease secreted by Bacillus clausii of industrial significance. METHODS AND RESULTS: Maximum enzyme activity was produced when the bacterium was grown in the medium containing (g l-1): soyabean meal, 15; wheat flour, 10; liquid maltose, 25; K2HPO4, 4; Na2HPO4, 1; MgSO4.7H2O, 0.1; Na2CO3, 6. The enzyme has an optimum pH of around 11 and optimum temperature of 60 degrees C. The alkaline protease showed extreme stability towards SDS and oxidizing agents, which retained its activity above 75 and 110% on treatment for 72 h with 5% SDS and 10% H2O2, respectively. Inhibition profile exhibited by phenylmethylsulphonyl fluoride suggested that the protease from B. clausii belongs to the family of serine proteases. CONCLUSIONS: Bacillus clausii produced high levels of an extracellular protease having high stability towards SDS and H2O2. SIGNIFICANCE AND IMPACT OF THE STUDY: The alkaline protease from B. clausii I-52 is significant for an industrial perspective because of its ability to function in broad pH and temperature ranges in addition to its tolerance and stability in presence of an anionic surfactant, like SDS and oxidants like peroxides and perborates. The enzymatic properties of the protease also suggest its suitable application as additive in detergent formulations.     

Purification and characterization of extracellular alkaline serine protease from Stenotrophomonas maltophilia strain S-1

AIMS: The present study was conducted by screening soil bacteria in an attempt to isolate a bacterium that produced extracellular alkaline protease, and for purification and characterization of the protease. METHODS AND RESULTS: Soil bacteria were screened by growth on casein as the sole carbon source. Characterization of a strain isolated from soil of Abashiri, Japan indicated a taxonomic affiliation to Stenotrophomonas maltophilia, and was named S-1 strain. The purified S-1 protease, designed S. maltophilia Protease-1 (SmP-1), exhibited an optimal pH of 12.0, optimal reaction temperature of 50 degrees C and a molecular mass of approximately 40 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The cleavage sites of the oxidized-insulin B chain by SmP-1 were identified as Leu6-Cys7, Cys7-Gly8, Tyr16-Leu17 and Leu17-Val18. The N-terminal amino acid sequence of the purified alkaline protease was determined as NH2-SASAPMVSGVAALVLE. CONCLUSION: A novel extracellular alkaline serine protease was isolated from S. maltophilia strain S-1. The optimal pH of the proteolytic activity was pH 12.0. SIGNIFICANCE AND IMPACT OF THE STUDY: The extremely high optimal pH and heat stability of the alkaline serine protease SmP-1 might make it widely applicable to food and other industries.