Microbial nitrilases have attracted increasing attention in nitrile hydrolysis for carboxylic acid production in recent years. A bacterium with nitrilase activity was isolated and identified as Pseudomonas putida CGMCC3830 based on its morphology, physiological and biochemical characteristics, as well as 16S rRNA gene sequence. The nitrilase production was optimized by varying culture conditions using the one-factor-at-a-time method and response surface methodology. Glycerol 13.54 g/L, tryptone 11.59 g/L, yeast extract 5.21 g/L, KH2PO4 1 g/L, NaCl 1 g/L, urea 1 g/L, initial pH 6.0 and culture temperature 30 degrees C were proved to be the optimal culture conditions. It resulted in the maximal nitrilase production of 36.12 U/mL from 2.02 U/mL. Investigations on substrate specificity demonstrate P. putida nitrilase preferentially hydrolyze aromatic nitriles. When applied in nicotinic acid synthesis, 2 mg/mL P. putida cells completely hydrolyzed 20.8 g/L 3-cyanopyridine into nicotinic acid in 90 min. The results indicated P. putida CGMCC3830 displayed potential for industrial production of nicotinic acid.
Aminohydrolases ; biosynthesis ; Culture Media ; Hydrolysis ; Niacin ; biosynthesis ; Nitriles ; metabolism ; Pseudomonas putida ; enzymology ; Pyridines ; metabolism ; RNA, Ribosomal, 16S ; genetics ; Substrate Specificity ; Temperature

Objective To evaluate the efficacy of combination solifenacin and tamsulosin for the treatment of distal ureteral calculi after extracorporeal shock wave lithotripsy. Methods 120 patients (male:100 female:20 age:18-67 yrs) randomly assigned to 4 groups (each group 30) with the calculi diameter range from 0.5 to 1.1 cm.All patients performed extracorporeal shock wave lithotripsy (X ray oriented).The control group did not accept any medical treatment.The solifenacin group were administered solifenacin 5 mg,once per day.The tamsulosin group were administered tamsulosin 0.2 mg,once per day.The combination group were administered solifenacin 5mg,plus tamsulosin 0.2 mg,each per day.The observation duration was set at 2 weeks. Results The stone-free rate (according to KUB) within 2 weeks were 80.0％,83.3％,93.3％ and 96.7％ in the control group,solifenacin group,tamsulosin group and combination group respectively.Statistical differences were significant among the tamsulosin group,the combination group and the control group.The stone expulsion times were (7.6 + 3.7) d,(6.3 ± 2.5) d,(4.4 + 2.3) d and (3.5 ± 2.2) d in the 4 groups respectively.Statistical differences were significant among the tamsulosin group,the combination group and the control group.The uses of analgesics were 13,5,9 and 3 in the 4 groups respectively.The bladder irrtative symptoms were 12,6,4 and 4 in the 4 groups respectively.Statistical differences were also significant for the use of analgesics and relief of bladder irritation between the solifenacin group,the combination group and the control group. Conclusions Tamsulosin and solifenacin could be safe and effective for the treatment of distal ureteral calculi after extracorporeal shock wave lithotripsy.It could significantly improve the stone expulsion rate,relief the pain and improve bladder irrtative symptoms.

OBJECTIVETo study the relationship between targeting vector structure and homologous recombination rate and investigate whether the mouse p16(INK4a) plays a role in tumor suppression.

METHODSA conditional targeting vector with 2.0 kb EcoR I/Xba I fragment as short arm and 5.9 kb SpeI/NotI fragment as long arm was built. Of the 2 direct locus crossing- over(loxPs) in the vector, one was inserted at 240 bp upstream of the initiate code of p16(INK4a) exon 1a and the other at 1633 bp downstream of the initiate code. Both exon 1a and the selection marker Neo will be deleted in targeted cells when mediated by Cre. After linearlization and purification, t he targeting vector was introduced into ES cells through electroporation.

RESULTSTwenty-four G418- and gancyclovir-resistant ES cell colonies were picked out and one of them was confirmed as positive by Southern hybridization.

CONCLUSIONTargeting vectors with 2 TK genes flanking the homologous arms are likely to produce good result of homologous recombination.

Animals ; Anti-Bacterial Agents ; pharmacology ; Antiviral Agents ; pharmacology ; Base Sequence ; Cell Division ; drug effects ; genetics ; Cell Line ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Drug Resistance ; genetics ; Embryo, Mammalian ; cytology ; drug effects ; metabolism ; Exons ; genetics ; Ganciclovir ; pharmacology ; Genetic Vectors ; genetics ; Gentamicins ; pharmacology ; Mice ; Molecular Sequence Data ; Recombination, Genetic ; Stem Cells ; cytology ; drug effects ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection

OBJECTIVETo identify and characterize laryngeal cancer related novel genes located on chromosome 6q25.

METHODSElectric hybridization was performed in human genome database using EST (expression sequence tag) as probe. Novel genes were deduced by software from positive DNA clones and their cDNAs were amplified by RT-PCR using primers designed according to the sequence of the putative genes.

RESULTSA novel gene was cloned successfully. The full length of this gene was about 21 kb. It contained two exons and produced a 1006 bp transcript coding a protein with 235 amino acid residues. It's 5'flanking sequence contained two binding sites of oncoprotein c-Myc, thus it was named MTLC (c-Myc target from laryngeal cancer cells). Homologous assay showed that MTLC exhibited little overall homology to known human proteins but it exhibited good overall homology to mouse MT-MC1 protein with an identity of 78%. The primary structure of MTLC protein contained a nuclear location signal motif, but it did not have other conserved domains. The results of subcellular location experiment showed that MTLC expressed in nuclei of human hepatocellular carcinoma cell line Bel7402 cells, while a wide distribution of MTLC in various tissues was demonstrated by Northern blotting.

CONCLUSIONMTLC may play an important role as a target gene of c-Myc and as a transcription factor in keeping the normal physiological process of cells.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Chromosomes, Human, Pair 6 ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Gene Expression ; Green Fluorescent Proteins ; Humans ; Laryngeal Neoplasms ; genetics ; Luminescent Proteins ; genetics ; metabolism ; Microscopy, Fluorescence ; Molecular Sequence Data ; Nuclear Proteins ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Tumor Cells, Cultured

Next generation sequencing technology has revolutionized studies in fermentation process, in particular, to explore the mechanism by which food microorganisms, including physiology, metabolic pathways, diversity and dynamic changes of microbial community. In addition, phylogenetic characteristics of different species or strains of the food microorganisms are disclosed. All these aspects will help explain how the microbes are interacting and responding to environmental factors. Bioinformatics analysis of genome and metagenome sequence data of food microorganisms could provide essential clues to improve fermentation process and function of microbes as well as control and prevention of foodborne disease outbreak. In this review, we summarized recent genomics and metagenomics studies on food microorganisms. The impact of next generation sequencing for the development and trends of food microorganism researches were discussed in details.
Computational Biology ; Food Microbiology ; Genomics ; High-Throughput Nucleotide Sequencing ; Metabolic Networks and Pathways ; Metagenomics ; Phylogeny