十三烷1,13—二羧酸的发酵研究

热带假丝酵母(C.tropicalis)NP-6-126是经过紫外线和亚硝酸反复诱变筛选培育出来的,它是生产十三烷1,13一二羧酸(DC_(15)的优良生产突变株,尿素和硝酸钾浓度对其产酸有明显影响。尿素浓度在0.15％～0.21％范围内对产酸有利,尤其是0.18％最佳,浓度增大,明显抑制DC_(15)的产生和积累;硝酸钾的加入,也明显促进DC_(15)产量的提高,0.6％～0.9％硝酸钾浓度较合适。于16L自动控制罐发酵7d,DC_(15)达到130g/L,放大到2500L罐,在最佳条件下,连续5批,发酵6d,DC_(15)产量平均达到176g/L,正十五烷(nC_(15)转化率平均为52.5％,后处理总收率平均为80.6％,DC_(15)的纯度平均为95.83％。     

十三烷1,13-二羧酸的发酵研究

热带假丝酵母(C.tropicalis)NP-6-126是经过紫外线和亚硝酸反复诱变筛选培育出来的,它是生产十三烷1,13一二羧酸(DC_(15)的优良生产突变株,尿素和硝酸钾浓度对其产酸有明显影响。尿素浓度在0.15％～0.21％范围内对产酸有利,尤其是0.18％最佳,浓度增大,明显抑制DC_(15)的产生和积累;硝酸钾的加入,也明显促进DC_(15)产量的提高,0.6％～0.9％硝酸钾浓度较合适。于16L自动控制罐发酵7d,DC_(15)达到130g/L,放大到2500L罐,在最佳条件下,连续5批,发酵6d,DC_(15)产量平均达到176g/L,正十五烷(nC_(15)转化率平均为52.5％,后处理总收率平均为80.6％,DC_(15)的纯度平均为95.83％。     

丙烯酸对十六碳二元酸发酵的影响和16L罐扩试

本文报道丙烯酸对发酵生产十六碳二元酸(DC_(16))的影响,加入0.1%丙烯酸,DC_(16)产量提高20—30%.在16L自动控制罐上,在最佳条件下,加入20%(v/v)正十六烷(nC_(16)),发酵5天,DC_(16)高达120g/L,nC_(16)的转化率高达79%.     

发酵生产十六烷二羧酸的研究

以热带假丝酵母(Candida tropicalis)T_25-14为出发菌株,经紫外线多次诱变,获得生产十六烷二羧酸(DC₁₆)能力比原株提高25％以上的4株突变株。其中UH-3-9突变株经多次复筛,产DC16能力都比T25-14,菌株提高50％以上。经摇瓶条件试验,不加其他生长碳源。只加15％(v／v)正十六烷(nCl6),发酵96h,DC16为48．2g/L,转化率41％,产品纯度95．9％。     

微生物发酵生产十一烷1，1 1二羧酸的研究

从热带假丝酵母(Candiada tropicalis)T25—14经过紫外线和亚硝酸的多次诱变,获得4株产十一烷l,11二羧酸(DC₁₃)较多的突变株,其中最优的NP-159株以20％(V／V)正十三烷(nC₁₃)为碳源摇瓶发酵4天,DC₁₃达80g／L左右。在16L罐上,以30％(V／V)nC₁₃发酵6天,DC₁₃高达139g／L,回收残烃后,对nC₁₃的转化率为80％以上。后处理收率为78．9％,DC₁₃的纯度为95．3％。     

酿酒酵母by1.1b产D-(-)-扁桃酸脱氢酶的发酵条件优化

对一株产D-(-)-扁桃酸对映选择性脱氢酶的酿酒酵母菌(Saccharomyces cerevisiae sp. strain by1.1b)发酵产酶条件进行了优化。研究各种碳源、氮源及无机盐对产酶的影响, 应用正交试验优化发酵培养基组成, 结果为: 蛋白胨 60 g/L, 麦芽糖 30 g/L, MgSO4 0.5 g/L, ZnSO4 0.01 g/L, KCl 1.0 g/L。优化后酶产量提高了7.9倍(由2.56 U/mL增至20.21 U/mL)。摇瓶培养最佳条件为: 装液量40 %, 发酵pH 6.5, 接种量10 %, 发酵温度30 ℃。考察了细胞生长及产酶的时间进程, 最佳培养时间为25 h。     

酿酒酵母by1.1b产D-(-)-扁桃酸脱氢酶的发酵条件优化

对一株产D-(-)-扁桃酸对映选择性脱氢酶的酿酒酵母菌(Saccharomyces cerevisiae sp.strain by1.1b)发酵产酶条件进行了优化.研究各种碳源、氮源及无机盐对产酶的影响,应用正交试验优化发酵培养基组成,结果为:蛋白胨60 g/L,麦芽糖30 g/L,MgSO4 0.5 g/L,ZnSO4 0.01 g/L,KCl 1.0 g/L.优化后酶产量提高了7.9倍(由2.56 U/mL增至20.21 U/mL).摇瓶培养最佳条件为:装液量40%,发酵pH 6.5,接种量10%,发酵温度30℃.考察了细胞生长及产酶的时间进程,最佳培养时间为25 h.     

红酵母类胡萝卜素高产菌株的筛选及其发酵生理学条件研究

从数株红酵母中选出了1株产类胡萝卜素能力较强的红酵母RY－98（生物量,类胡萝卜素含量和产量分别为19．9g/L,334.8ug/g和6．7mg/L);研究了该菌株产类胡萝卜素的最适营养与环境条件,获得了最佳的发酵生理学条件;葡萄糖40g/L,(NH4)2SO4 10g/L,酵母膏3g/L,蕃茄汁2mL/L,花生油0.5mL/L,接种量30mLL初始pH6.0和通气量（培养基装置040ml/250mL:,在此初步优化的培养条件下,红酵母RY－98经72小时摇瓶发酵其生物量,类胡萝卜素含量和产量分别可达26．8g/L,386.9ug/g和10．4mg/L,依次比初筛中提高了34．7％,15．6％,和55．2％。     

红酵母类胡萝卜素形成的生理学研究

从数株红酵母中选出 1株产类胡萝卜素能力较强的红酵母RY 98(生物量、类胡萝卜素含量和产量分别为19.9g/L ,334 .8μg/ g和 6 .7mg/L) ;研究了该菌株产类胡萝卜素的最适营养与环境条件 ,获得了最佳的发酵生理学条件 :葡萄糖 40 g/L ,(NH4 ) 2 SO4 10 g/L ,酵母膏 3g/L ,蕃茄汁 2mL/L ,花生油 0 .5mL/L ,接种量 30mL/L ,初始pH 6 .0和通气量 (培养基装量 ) 4 0mL/ 2 5 0mL。在此初步优化的培养条件下 ,红酵母RY 98经 72h摇瓶发酵其生物量、类胡萝卜素含量和产量分别可达 2 6 .8g/L ,386 .9μg/ g和 10 .4mg/L ,依次比初筛中提高了 34 .7% ,15 .6 %和 5 5 .2 %。     

1,3-丙二醇产生菌的诱变育种及培养基优化

以实验室自然筛选的克雷伯氏杆菌(Klebsiella sp.)为出发株,采用紫外诱变及亚硝基胍和超声波协同处理获得一株1,3-丙二醇高产突变株。在摇瓶发酵中,其产1,3-丙二醇产量由17.39 g/L提高到24.11 g/L,提高38.64%。变异株经10次传代培养,发酵能力稳定。对发酵培养基成分进行了优化,优化后1,3-丙二醇产量为30.05g/L,为优化前的1.25倍。     

Ring (15) chromosome

Summary Cytogenetic studies on lymphocytes from a girl aged 3 years and 10 months revealed a ring chromosome 15. Several banding methods showed the r(15) chromosome not to have any apparent deletion of the long arm. The silver staining technique for nucleolar organizer regions showed an NOR positive region (band p12). In only a few cells was a chromosome 15 missing. The size of the r(15) was found to be constant. Comparison with 11 previous reported cases in the literature shows that the clinical manifestations in the different patients with ring chromosome 15 are constant although not clinically identifiable and it appears likely to attribute them to a significantly retarded intrauterine and postnatal growth instead of presumed deficiency in the long arm and mosaic configurations.     

15-hydroxyprostaglandin dehydrogenase (15-PGDH) and lung cancer

15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes NAD(+)-linked oxidation of 15 (S)-hydroxyl group of prostaglandins and lipoxins and is the key enzyme responsible for the biological inactivation of these eicosanoids. The enzyme was found to be under-expressed as opposed to cyclooxygenase-2 (COX-2) being over-expressed in lung and other tumors. A549 human lung adenocarcinoma cells were used as a model system to study the role of 15-PGDH in lung tumorigenesis. Up-regulation of COX-2 expression by pro-inflammatory cytokines in A549 cells was accompanied by a down-regulation of 15-PGDH expression. Over-expression of COX-2 but not COX-1 by adenoviral-mediated approach also attenuated 15-PGDH expression. Similarly, over-expression of 15-PGDH by the same strategy inhibited IL-1beta-induced COX-2 expression. It appears that the expression of COX-2 and 15-PGDH is regulated reciprocally. Adenoviral-mediated transient over-expression of 15-PGDH in A549 cells resulted in apoptosis. Xenograft studies in nude mice also showed tumor suppression with cells transiently over-expressing 15-PGDH. However, cells stably over-expressing 15-PGDH generated tumors faster than those control cells. Examination of different clones of A549 cells stably expressing different levels of 15-PGDH indicated that the levels of 15-PGDH expression correlated positively with those of mesenchymal markers, and negatively with those of epithelial markers. It appears that the stable expression of 15-PGDH induces epithelial-mesenchymal transition (EMT) which may account for the tumor promotion in xenograft studies. A number of anti-cancer agents, such as transforming growth factor-beta1 (TGF-beta1), glucocorticoids and some histone deacetylase inhibitors were found to induce 15-PGDH expression. These results suggest that tumor suppressive action of these agents may, in part, be related to their ability to induce 15-PGDH expression.     

Gastric antisecretory effect of 15(R)-15-menthyl PGE2, methyl ester and of 15(S)-15-methyl ester.

Gastric juice was collected from gastric pouches in dogs stimulated with histamine. 15(R)-15-methyl PGE2, methyl ester inhibited gastric secretion in dogs when given orally, but was almost inactive when given intravenously, whereas 15(S)-15-methyl PGE2 methyl ester was active by both routes. When given directly into the small intestine (intrajejunally), the 15(S) was active and the 15(R) was inactive. The 15(R), diluted in acid and administered intrajejunally, became active in inhibiting gastric secretion. When the 15(S) was diluted in acid and administered intrajejunally, it lost half of its activity. When each analog was incubated in an acid medium, each was epimerized to give approximately a 1:1 mixture of both 15(R) and 15(S). Incubation of the 15(R) in pH 3 buffer resulted in only a trace of formation of 15(S). These results explain why the 15(R) is active orally but not intrajejunally. When given orally, the low pH of gastric secretion epimerizes much of the 15(R) into the 15(S),which is active by any route. The degree of acidity of gastric contents may determine whether the 15(R) will exert an antisecretory effect.     

The purine-rich trinucleotide repeat sequences d(CAG)15 and d(GAC)15 form hairpins.

The structures of single-stranded (ss) oligonucleotides containing (CAG)15 [ss(CAG)15] or (GAC)15 [ss(GAC)15] were examined. At 10 degrees C, the electrophoretic mobilites of the two DNAs were similar to ss(CTG)15, a DNA that forms a hairpin containing base paired and/or stacked thymines. At 37 degrees C in 50 mM NaCl, single-strand-specific P1 nuclease cleaved the G33-G36 phosphodiesters of ss(GAC)15, and the G32-A34, G35-C36 phosphodiesters of ss(CAG)15 (where the loop apex of both DNAs = A34). Electrophoretic mobility melting profiles indicated that the melting temperature (Tm) of ss(CAG)15 in low (approximately 1 mM Na+) ionic strength was 38 degrees C. In contrast, the Tm of ss(GAC)15 was 49 degrees C, a value similar to the Tm of ss(CTG)15. These results provide evidence that ss(GAC)15 and ss(CAG)15 form similar, but distinguishable hairpin structures.     

The Eps15 homology (EH) domain

The Eps15 homology (EH) domain was originally identified as a motif present in three copies at the NH2-termini of Eps15 and of the related molecule Eps15R. Both of these molecules are substrates for the tyrosine kinase activity of the epidermal growth factor receptor and hence the name 'Eps15 homology' or EH domain [Wong et al. (1994) Oncogene 9, 1591-1597; Wong et al. (1995) Proc. Natl. Acad. Sci. USA 92, 9530-9534; Fazioli et al. (1993) Mol. Cell. Biol. 13, 5814-5828] was derived. The motif was subsequently found in several proteins from yeast to nematode, thus establishing its evolutionary conservation. Initial studies with filter-binding assays and phage-displayed libraries demonstrated its protein:protein interaction abilities and identified specific ligands. Subsequently, structural analyses established the molecular bases of recognition between EH domains and cognate peptides. To date, several EH-containing and EH-binding proteins have been identified, which establish in the cell a network of protein:protein interactions, defined as the EH network. This network coordinates cellular functions connected with endocytosis, actin remodeling and intracellular transduction of signals.     

Transmission of stability (telestability) in deoxyribonucleic acid. Physical and enzymatic studies on the duplex block polymer d(C15A15) - d(T15G15).

The properties of the duplex block polymer d(C15A15) - d(T15G15) were examined by thermal denaturation and nuclease susceptibility studies in the absence and presence of drugs (actinomycin and netropsin) which bind specifically to only one end of the block polymer. The nucleotide composition of one region of this synthetic double-helical DNA affected the properties of a contiguous but remote region. Furthermore, the binding of actinomycin influenced the properties of both the binding and nonbinding regions. These findings suggest a mechanism for gene regulation at a distance.