A Convenient Method for the Quantitative Determination of Pipecolic Acid

The quantitative determination of pipecolic acid was examined.

The reaction of 3% ninhydrin solution in n-butanol, saturated with citrate buffer (pH 4.2), with pipecolic acid in boiling water for 3 min yielded the colored products showing λ_max at 570 mμ, but with proline hardly yielded those products. By the colorimetry proposed, it is possible to determine the amount of pipecolic acid in the sample containing proline no more than 50 times the amount of the pipecolic acid, directly from the calibration curve using pipecolic acid.

The method for removal of amino acids from the sample containing pipecolic acid and proline was examined and discussed.     

31P NMR Spectra of Alkalophilic Bacillus No. A-59

Some advanced cancer patients suffer from pungent sulfury malodor. To determine the chemical identity of the odorant, we performed gas chromatography-mass spectrometry-olfactometry analysis of volatiles from fungating cancer wounds. We identified the source of the characteristic smell as dimethyl trisulfide, a compound that is known to be emitted from some vegetables and microorganisms. Controlling the production of dimethyl trisulfide should improve quality of life of patients.     

Transfructosylation of Thiol Group by β-Fructofuranosidases

β-Fructofuranosidase fructosylated not only the hydroxyl group but also the thiol group of 2-mercaptoethanol in a transfer reaction using sucrose as a donor substrate. The enzymes from Candida utilis and Saccharomyces cerevisiae (bakers’ yeast) were effective catalysts for the thio-fructofuranosylation. The thio-fructosylation product was isolated by activated carbon chromatography and its structure was confirmed by Fab-mass spectrometry and NMR spectroscopy. The thio-fructofuranoside was synthesized effectively at around 3.0 M for the acceptor concentration. The product increased with the sucrose concentration at least up to 1.9 M. O-Fructofuranoside was simultaneously synthesized at an early stage of the reaction, although it was hydrolyzed on further incubation. On the contrary, the thio-furctofuranoside accumulated efficiently after synthesis, indicating it was very stable against the hydrolytic action of the β-fructofranosidase.     

Chemical Identity of a Rotting Animal-Like Odor Emitted from the Inflorescence of the Titan Arum (Amorphophallus titanum)

The titan arum, Amorphophallus titanum, is a flowering plant with the largest inflorescence in the world. The flower emits a unique rotting animal-like odor that attracts insects for pollination. To determine the chemical identity of this characteristic odor, we performed gas chromatography-mass spectrometry-olfactometry analysis of volatiles derived from the inflorescence. The main odorant causing the smell during the flower-opening phase was identified as dimethyl trisulfide, a compound with a sulfury odor that has been found to be emitted from some vegetables, microorganisms, and cancerous wounds.     

Amino Acid Composition of Cucumber Green Mottle Mosaic Virus Coat Protein and Sequence Determination of Its Tryptic Peptides

Amino acid composition of the CGMMV* coat protein was determined to be as follows: Asp₂₀, Thr₁₀, Ser₂₄, Glu₁₀. Pro₆, Gly₉, Ala₂₁, Val₇, Ile₇, Leu₁₈, Tyr₄, Phe₉, Lys₄, His₁, Arg₈, Trp₂. No terminal α-amino group was detected by dinitrophenylation method. The carboxyl-terminus was found to be serine by hydrazinolysis of the protein and digestion with carboxypeptidase A.

For sequence analysis of the coat protein, tryptic digestion was accomplished at pH 8.0 resulting in ten soluble and several insoluble peptides at pH 4.5. The amino acids contained in soluble peptides accounted for 91 out of 160 residues in the whole protein. The amino acid sequences of ten soluble peptides were determined.

From the similarities of amino acid sequence of the peptides to those of TMV* protein, CGMMV was assumed to be a strain of TMV group.     

鲍曼不动杆菌耐消毒剂基因检测

目的 了解深圳市人医院鲍曼不动杆菌抗季铵盐类消毒剂基因的检出率及基因型特点.方法 收集深圳市人民医院临床标本中分离鲍曼不动杆菌67株,PCR检测分离株的qacA/B、qacE△1、qacC、qacG和qacJ 5种抗季铵盐类消毒剂基因.结果 67株鲍曼不动杆菌中qacE△l基因阳性42株(62.6％),qacA/B与qacG基因各有一菌株阳性,未检测出qacC或qacJ基因阳性菌株.结论 深圳市人医院鲍曼不动杆菌中抗季铵盐类消毒剂基因检出率较高,主要为qacE△1型.     

不依赖连接反应的高通量克隆方法

以脱脂菜籽粕酶解的复合氨基酸为主要原料,螯合率为考察指标,采用L16(4 5)正交试验设计,考察了pH值、温度、时间和配位比对螯合率的影响.结果显示,影响因素的高低顺序为:配位比＞pH值＞时间＞温度.结果表明,复合氨基酸与铜螯合的主要影响因素为pH值和配位比,且配位比的影响达到极显著水平.最佳工艺条件为:时间50 min,温度50℃,配位比2∶1,pH为9,此条件下的复合氨基酸螯合铜的螯合率为94.59％,氨基酸含量为30.2％.     

瓜蚜为害对寄主植物主要营养物质和次生物质的影响

为明确瓜蚜Aphis gossypii Glover与其寄主植物相互作用关系及其机理,本文采用蒽酮比色法、考马斯亮蓝法、磷钼酸－磷钨酸比色法及索氏回流法,分别测定了瓜蚜为害对黄瓜、哈密瓜、南瓜、瓢葫芦和搅瓜等5种寄主植物叶片内可溶性总糖、可溶性蛋白质、单宁及黄酮含量的影响。结果表明,瓜蚜为害后,供试寄主植物叶片中可溶性糖和可溶性蛋白含量显著降低,而单宁和黄酮的含量明显上升。说明瓜蚜为害导致寄主植物营养质量下降,而次生代谢物质含量上升,从而对植食者产生诱导抗性。     

微杆菌Sneb159杀线虫活性物质的分离与鉴定

【目的】有关Microbacterium maritypicum在植物线虫生物防治方面的研究较少,探究菌株M. maritypicum Sneb159的杀线虫活性,明确菌株发酵液中具有杀线虫活性的物质,为生物农药的开发提供理论依据。【方法】本研究检测了菌株Sneb159发酵液对大豆胞囊线虫二龄幼虫的触杀活性;并以触杀活性为追踪,采用有机溶剂萃取、硅胶柱层析及半制备高效液相色谱等技术对菌株Sneb159发酵滤液中的活性物质进行分离纯化;采用核磁共振波谱对纯化物进行结构鉴定。【结果】菌株Sneb159具有杀线虫活性,在24 h和48 h处理组中线虫的死亡率均显著高于对照组。从菌株Sneb159发酵滤液中分离得到杀线虫活性物质A6,经结构鉴定确定该物质为苯乙酰胺。【结论】首次发现M. maritypicum对大豆胞囊线虫的触杀作用,并且明确活性物质为苯乙酰胺。结果表明菌株M. maritypicum Sneb159和苯乙酰胺在大豆胞囊线虫的生物防治方面具有较好的应用潜力。