Biotransformation of podophyllotoxin to picropodophyllin by microbes

Biotransformation of podophyllotoxin (PT) by several microbial species has been investigated. Among the fungi tested, it was found that Penicillium strains can isomerize PT to picropodophyllin (PPT) in 8% yield and other strains also transform the substrate into the same product but with lower yield.     

A new p-hydroxybenzoic acid derivative from an endophytic fungus Penicillium sp. of Nerium indicum

A new p-hydroxybenzoic acid derivative named 4-(2′R, 4′-dihydroxybutoxy) benzoic acid (1) was isolated from the fermentation of Penicillium sp. R22 in Nerium indicum. The structure was elucidated by means of spectroscopic (HR-ESI-MS, NMR, IR, UV) and X-ray crystallographic methods. The antibacterial and antifungal activity of compound 1 was tested, and the results showed that compound 1 revealed potent antifungal activity against Colletotrichum gloeosporioides, Alternaria alternata, and Alteranria brassicae with MIC value of 31.2 μg/ml.     

南极真菌Penicillium sp. S-2-10次级代谢产物的研究

目的 对来源于南极海域海底沉积物（47.09° W,62.05° S,水深1393 m）真菌Penicillium sp. S-2-10的次级代谢产物进行研究。方法 对菌株进行规模发酵,并对所得发酵液的乙酸乙酯萃取浸膏进行系统分离纯化,据化学指征和生物学活性两方面特点,采用Sephadex LH 20凝胶色谱、正相硅胶柱色谱、反相ODS柱色谱、高效液相柱层析等色谱分析方法,通过核磁（1H-NMR和13C-NMR等）及质谱（MS）等波谱分析方法,并与相关文献比较,鉴定化合物结构。 结果 分离得到7个单体化合物,经鉴定化合物1~7分别为Benzoic acid, 2-(2,6-dihydroxybenzoyl)-3-hydroxy-5-(hydroxymethyl)-methyl, methyl ester (1), chrysophanol (2), Volemolide (3), Ergosta-6,8(14),22-trien-3-ol (4), 2-aminophenoxazin-3-one (5), 2-benzothiazolol (6), benzoic acid (7)。结论 化合物3,4为首次从Penicillium属真菌中获得。     

Differences in spontaneous mutation frequencies as a function of environmental stress in soil fungi at "Evolution Canyon," Israel

When various wild strains of Penicillium lanosum and Aspergillus niger were placed in the same mild laboratory environment, their frequencies of new spontaneous mutations were clearly related to whether they had been isolated from a region of high or low microclimatic stress. In the mild environment, the total frequencies of conidial color and morphological mutations in P. lanosum, summed over all relevant loci, ranged from 0.29% to 2.4% for six strains from the north-facing, less stressful "European" slope (ES/NFS) of "Evolution Canyon" I, compared with 6.5-11.6% for five strains from the south-facing "African" slope (AS/SFS), which is a much more stressful environment, being harsher, drier, more fluctuating in temperature, and receiving up to eight times more UV radiation than the opposite slope. The corresponding figures for A. niger were 0.42-1.50% for three strains from the ES/NFS and 2.3-4.9% for six strains from the AS/SFS. The more mutagenic environment of the AS/SFS than of the ES/NFS means that, in Evolution Canyon, the mutation frequency differences between the very stressful environment and the less stressful environment are probably even larger than the 4- and 6-fold differences found here in a mild laboratory environment. The evidence from these two filamentous fungi, which have no sexual cycle, is that there are inherited differences in spontaneous mutation rates according to the levels of stress in the environment, and this feature may well be adaptive. Evolution Canyon I is at Nahal Oren, Mount Carmel, Israel.     

南极真菌Penicillium sp. S-3-88次级代谢产物的研究

目的 对源于南极(44.42° W,60.54° S,水深239 m,水温-1.16 ℃)海参体内共附生真菌Penicillium sp. S-3-88的次级代谢产物进行研究。方法 采用Sephadex LH-20色谱、反相ODS柱色谱、高效液相柱层析等色谱分析方法,对Penicillium sp. S-3-88发酵液的乙酸乙酯萃取部位进行系统分离,并通过₁H和₁₃C NMR及质谱(LC-MS)等现代波谱解析手段,并与相关文献比对,鉴定化合物结构。结果 分离得到9个单体化合物。结论 鉴定化合物 1 – 9 分别为2-acetyl-4(3H)-quinazolin--one(1), benzo[d]thiazol-2(3H)-one(2), Octadecyl phenylpropanoate(3), diisobutyl phthalate(4), butyl-isobutyl-phthalate(BIP)(5), ergosterol peroxide(6), melithasterol B(7),22E,24R-5α,6α-epoxyergosta-8(14),22-diene-3β,7α-diol(8)和cerevisterol(9)。     

Purification of a potent mitogenic homodimeric Penicillium griseoroseum lectin and its characterisation

Penicillium griseoroseum lectin was 80‐fold purified by successive DEAE Sepharose anion exchange and Sephadex G‐100 gel permeation chromatography. P. griseoroseum lectin exhibited haemagglutination activity towards protease‐treated rabbit erythrocytes. It showed specificity towards various carbohydrates such as d ‐mannose, N‐acetyl‐d ‐glucosamine, mucins, and so forth. P. griseoroseum lectin was found as a glycoprotein with glycan content of 4.33%. Purified P. griseoroseum lectin is homodimeric having a molecular mass of 57 kDa with subunit molecular mass of 28.6 kDa. Haemagglutination activity of purified P. griseoroseum lectin was completely stable from 25°C to 35°C at a pH range of 6–7.5. Lectin activity was not influenced by divalent metal ions and denaturants. P. griseoroseum lectin manifested mitogenicity towards mice splenocytes and activity reached a peak at 75 μg/ml of lectin concentration. P. griseoroseum lectin in microgram concentrations stimulated proliferation of mice splenocytes. Thus, P. griseoroseum lectin exhibits potential mitogenicity, which can be exploited for further biomedical applications.     

一株提高刺五加苷E含量内生真菌的鉴定及其作用机制初探

目的确定分离自刺五加的内生真菌P116-1a的分类地位,并初步明确其提高刺五加苷E含量的作用机制。方法应用形态学及ITS序列分析方法进行鉴定,将菌液回接刺五加浸出液进行发酵培养和将灭活后的菌液注射刺五加后,HPLC法分析刺五加苷含量的变化。结果 P116-1a与Penicillium minioluteum的形态学特征相符,且与其ITS序列同源性最高(98.6%)。在培养早期,活体P116-1a可显著提高刺五加浸出液中刺五加苷E的含量,灭活后其对刺五加苷E含量无显著作用。结论 P116-1a为Penicillium minioluteum,其作用方式为通过调节刺五加苷生物合成关键酶的表达、酶蛋白的活性而提高刺五加苷E的含量。     

巢式PCR检测石蜡包埋组织中着色霉、孢子丝菌和马尔尼菲青霉

目的 评价巢式PCR对不同深部真菌感染组织石蜡包埋标本检测的价值。方法 收集着色芽生菌病、孢子丝菌病、马尔尼菲青霉病及其他深部真菌病石蜡组织标本共44份,行组织病理观察并提取石蜡组织标本中DNA。使用针对着色霉、孢子丝菌及马尔尼菲青霉核糖体DNA特定区域的特异性巢式PCR引物,分别对所提取的真菌DNA进行扩增。分析巢式PCR对这3种病原真菌扩增的敏感性和特异性,并与组织病理检查方法比较。结果 20例着色芽生菌病组织蜡块中8例扩增阳性,10例孢子丝菌病组织蜡块中7例扩增阳性,10例马尔尼菲青霉病组织蜡块均扩增阳性,其余对照深部真菌病组织蜡块扩增均为阴性,巢式PCR检测3种真菌的敏感性分别为40%、70%和100%,特异性均达到100%。组织病理检查3种真菌的阳性率分别为95%、70%、80%。结论 巢式PCR扩增石蜡包埋组织中的真菌DNA是诊断深部真菌病的一种方法,尤其适用于诊断马尔尼菲青霉病。     

艾滋病患者马尔尼菲青霉菌优势株酵母相全长cDNA文库的构建和初步分析

Objective To construct a full length cDNA library of the dominant strain of Penicillium marneffei (PM) in yeast phase isolated from AIDS patients in Guangdong province and screen UniGenes as well as full-length genes, so as to establish the foundation for the study of PM's functional genes and pathogenic mechanisms. Methods CloneMiner cDNA construction kit was utilized to extract mRNA of the dominant PM strain isolated from AIDS patients in Guangdong province. The mRNA was reversed into cDNA, then cloned into a pDONR222 vector by BP recombination to obtain an Uncut cDNA library, which was homogenized later to construct a normalized cDNA library with the principal of saturation hybridization for DNA genome. 2000 clones were chosen randomly to make a bi-directional sequencing and analyzed with bioinformatics for screening UniGenes and full-length genes. Results The total clone number of the Uncut cDNA library was 1.16 × 107 cfu/mL, with a recombination rate of 95% and an average insertion element being over 1 kb. The total clone number of the normalized cDNA library was 1.18 × 106 cfu/mL, with a recombination rate of 95% and an average insertion element being over 1 kb as well. 1945 genes which DNA length were longer than 1 kb were obtained by sequencing and merged into 1360 UniG enes, of which 632 genes were full-length ones. Conclusions The full-length cDNA library of the dominant strain of PM from AIDS patients in Guangdong province possesses good quality.Meanwhile, the technical routine presents high efficiency in obtaining full-length genes and establishing a gene expression spectrum, which can contentedly meet the needs of future experiments.