Statistical Optimization of Culture Media and Conditions for Production of Mannan by Saccharomyces cerevisiae

Hong-Zhi Liu,Qiang Wang,Yuan-Yuan Liu,Fang Fang 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.5

In view of the increase in Saccharomyces cerevisiae mannan content, the culture medium and condition for S.cerevisiae were optimized in this study. The influence of culture medium ingredients such as carbon and nitrogen sources, inorganic ion, and enzyme activator on mannan production were evaluated using factional design. The mathematical model was established by the quadratic rotary combination design through response surface analysis. The optimized concentrations of culture medium were determined as follows: 4.98 g/100 mL, sucrose; 4.39 g/100 mL, soybean peptone; 3.10 g/100 mL, yeast extract; and 2.21 g/100 mL, glycerol. The optimized culture medium increased mannan production from 82.7 ± 3.4 mg/100 mL to 162.53 ± 3.47 mg/100 mL. The influence of original pH, inoculum size, temperature, and media volume on mannan production was evaluated and confirmed by orthogonale experimental design, with the order of effect as follows: media volume > temperature > initial pH > inoculation size. The optimized culture condition was pH, 5; inoculum size, 5 ml; temperature, 32oC; and media volume, 40 mL. The maximum mannan production increased to 258.5 ± 9.1 mg/100 mL at the optimum culture condition.=It was evident that the mannan production was affected significantly by culture medium and condition optimization (é < 0.01). In view of the increase in Saccharomyces cerevisiae mannan content, the culture medium and condition for S.cerevisiae were optimized in this study. The influence of culture medium ingredients such as carbon and nitrogen sources, inorganic ion, and enzyme activator on mannan production were evaluated using factional design. The mathematical model was established by the quadratic rotary combination design through response surface analysis. The optimized concentrations of culture medium were determined as follows: 4.98 g/100 mL, sucrose; 4.39 g/100 mL, soybean peptone; 3.10 g/100 mL, yeast extract; and 2.21 g/100 mL, glycerol. The optimized culture medium increased mannan production from 82.7 ± 3.4 mg/100 mL to 162.53 ± 3.47 mg/100 mL. The influence of original pH, inoculum size, temperature, and media volume on mannan production was evaluated and confirmed by orthogonale experimental design, with the order of effect as follows: media volume > temperature > initial pH > inoculation size. The optimized culture condition was pH, 5; inoculum size, 5 ml; temperature, 32oC; and media volume, 40 mL. The maximum mannan production increased to 258.5 ± 9.1 mg/100 mL at the optimum culture condition.=It was evident that the mannan production was affected significantly by culture medium and condition optimization (é < 0.01).

Use of In Vivo-Induced Antigen Technology to Identify In Vivo-Expressed Genes of Campylobacter jejuni During Human Infection

( Yuan Qing Hu ),( Jin Lin Huang ),( Qiu Chun Li ),( Yu Wei Shang ),( Fang Zhe Ren ),( Yang Jiao ),( Zhi Cheng Liu ),( Zhi Ming Pan ),( Xin An Jiao ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.3

Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.

Effects of Insect Hormones on the Replication of Nucleopolyhedrovirus

Zhang, Zhi-Fang,Yi, Yong-Zhu,Xiao, Qing-Li,He, Jia-Lu,Zhou, Ya-Jing,Zhang, Yuan-Xing Korean Society of Sericultural Science 2002 International Journal of Industrial Entomology Vol.4 No.2

An experimental study was undertaken to quantify the effects of insect hormones on the replication of nucleopolyhedrovirus (NPV). The results demonstrated that TCID/ sub 50/ at 72 h post-infection (hpi) rose systematically from 0.55$\times$10$^{8}$ /m1, for untreated cells, up to 1.67$\times$10$^{8}$ / ml at 3$\mu$g/ml, then dropped down to 1.45$\times$10$^{8}$ /m1 at 4 $\mu$g/ml, by adding ecdysone to the culture medium for Bm-N cells infected with a wild-type Bambyx mori. nucleopolyhedrovirus (BmNPV). The optimum enhancement of about 3 times on budded virus (BV) titer at 72 hpi was given at 3 $\mu$g/ml of ecdysone. While the polyhedra number had no obvious variation within the range of concentrations from 0 to 4 $\mu$g/ml. By addition of juvenile hormone analogue (JHA) into the media with this concentration range, the BmNPV TCID/ sub 50/ and polyhedra number at 72 hpi did not show significant changes. Also, the addition of either 3 $\mu$g/ml of ecdysone or 3 $\mu$g/ml of JHA to the culture media did not appear to affect the TCID/ sub 50/ and polyhedra number significantly in infected Sf-21 cells with the autographa californica nucleopolyhedrovirus (AcMNPV).

Antioxidant Peptide Purified from Enzymatic Hydrolysates of Isochrysis Zhanjiangensis and Its Protective Effect against Ethanol Induced Oxidative Stress of HepG2 Cells

Mei-Fang Chen,Yuan Yuan Zhang,Mei Di He,Cheng Yong Li,Chun Xia Zhou,Peng Zhi Hong,Zhong-Ji Qian 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.2

Marine microalgae have been widely applied in cosmaceuticals, nutraceuticals, and functional foods. In the present study, we first investigated the hepatoprotective effects of peptide purified from microalgae, Isochrysis zhanjiangensis on HepG2 cells alcoholic injury. I. zhanjiangensis was hydrolyzed utilizing chymotrypsin, trypsin, pepsin, and by vitro gastrointestinal digestion. Among hydrolysates, the gastrointestinal hydrolysate showed relatively high free radical scavenging ability preliminarily and was purified with following sequential chromatography methods. The amino acid sequence and molecular mass of the purified peptide from I. zhanjiangensis (PIZ) was identified as Asn-Asp-Ala-Glu-Tyr-Gly-Ile-Cys-Gly-Phe (NDAEYGICGF; MW, 1088.16 Da) via Q-TOF ESI/MS. Additionally, PIZ attenuated ethanol-induced cytotoxicity and inhibited the production of intracellular reactive oxygen species by DCFH-DA assay. Western blot results showed that superoxide dismutase (SOD) and glutathione (GSH) levels up-regulated with PIZ treatment before alcohol exposure while gamma-glutamyltransferase (GGT) protein expression down-regulated. These results provide an opportunity to discover new highly active peptide against alcohol toxicity in HepG2 cells.

Synthesis and characterization of monodisperse yttrium aluminum garnet (YAG) micro-crystals with rhombic dodecahedron

Xu, Tao,Yuan, Rui,Xu, Peng-Cheng,Pan, Dong-Jie,Yang, Woochul,Guo, Hai-Bo,Shen, Yu-Fang,Hu, Jian-Feng,Zhang, Zhi-Jun,Zhao, Jing-Tai Elsevier 2018 Journal of alloys and compounds Vol.762 No.-

<P><B>Abstract</B></P> <P>Uniform, monodisperse yttrium aluminum garnet (YAG) rhombic dodecahedron micro-crystals were synthesized using coprecipitation and hydrothermal methods. The Na<SUB>2</SUB>SO<SUB>4</SUB> as a surfactant was helpful in improving the dispersity of the crystallites and forming well-faceted, micro-sized dodecahedral YAG crystallites. The effect of the reaction time and the solvent has also been studied. The crystal growth mechanism called dissolution/crystallization was determined by analyzing the experimental and simulated results. And the specific well-developed crystal facets in the {110} family have been demonstrated by the theoretical calculation. A higher integrated emission intensity can be achieved for the YAG:Ce crystallites by improving the phase purity, morphology with better developed dodecahedrons, and dispersion. Furthermore, a rather high luminous efficacy (LE = 104.14 lm/W) for the white LED fabricated using the YAG:Ce<SUP>3+</SUP> phosphor prepared in this work was realized. The correlation between the crystal growth, morphology and luminescence properties of YAG crystallites has been established, which exhibits a great importance for the synthesis and application of mono-dispersed YAG crystallites.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Uniform and monodisperse YAG rhombic dodecahedron micro-crystals were synthesized. </LI> <LI> The growth mechanism of YAG rhombic dodecahedron was studied through experimental and computational simulation. </LI> <LI> The correlation between the crystal growth, morphology and luminescence of YAG crystallites was established. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

DNMT3a rs1550117 Polymorphism Association with Increased Risk of Helicobacter pylori Infection

Cao, Xue-Yuan,Jia, Zhi-Fang,Cao, Dong-Hui,Kong, Fei,Jin, Mei-Shan,Suo, Jian,Jiang, Jing Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

Background: DNA methyltransferase-3a (DNMT3a) plays significant roles in embryogenesis and the generation of aberrant methylation in carcinogenesis. This study aimed to investigate associations between single nucleotide polymorphisms (SNPs) of the DNMT3a gene and risk of Helicobacter pylori infection, gastric atrophy and gastric cancer. Methods: The subjects comprised 447 patients with gastric cancer; 111 individuals with gastric atrophy and 961 healthy controls. Two SNPs (rs1550117 and rs13420827) of the DNMT3a gene were genotyped by Taqman assay. DNMT3a expression was analyzed in cancer tissues from 89 patients by tissue microarray technique. Odds ratio (ORs) and 95% confidence intervals were calculated by multivariate logistic regression. Results: Among healthy controls, risk of H.pylori infection was significantly higher in subjects with the rs1550117 AA genotype, compared to those with GG/AG genotypes of DNMT3a [OR=2.08, (95%CI: 1.02-4.32)]. However, no significant correlation was found between the two SNPs and risk of developing gastric atrophy or gastric cancer. In addition, no increase in DNMT3a expression was observed in the gastric cancer with H.pylori infection. Conclusions: This study revealed that DNMT3a rs1550117 polymorphism is significantly associated with an increased risk of H. pylori infection, but did not support any evidence for contributions of DNMT3a rs1550117 and rs13420827 to either gastric atrophy or gastric cancer. The biological roles of DNMT3a polymorphisms require further investigation.

필자약력

거삼원(San Yuan Ju),공범군(Fan Jun Kong),김익수(Ick Soo Kim),박근(Keun Park),백진현(Jin Hyun Paik),송성유(Cheng You Song),오승열(Seung Yul Oh),장공자(Kong Ja Chang),조석흔(Xi Xin Zhao),진노직(Lu Zhi Chen),하방천(Fang Chuan He),한충민(C 한양대학교 아태지역연구센터 1995 중소연구 Vol.19 No.4

Mechanical behavior and chloride resistance of cementitious composites with PE and steel fiber

Liao, Qiao,Guo, Zhen-wen,Duan, Xin-zhi,Yu, Jiang-tao,Liu, Ke-ke,Dong, Fang-yuan Techno-Press 2021 Advances in concrete construction Vol.12 No.6

The mechanical behaviors and chloride resistance performance of fiber reinforced cementitious composites (FRCC) with hybrid polyethylene (PE) and steel fiber (in total 2% by volume) were investigated. Based on micro-mechanics and fracture mechanics, the reason why the tensile strain capacity of FRCC changed obviously was obtained. Besides, the effects of the total surface area of fiber in FRCC on compressive strength and chloride content were clarified. It is found that the improvement of the tensile strain capacity of FRCC with hybrid fiber is attributed to the growth of strain-hardening performance index (the ratio of complementary energy to crack tip toughness). As the total surface area of fiber related with the interfacial transition zone (ITZ) between fiber and matrix increases, compressive strength decreases obviously. Since the total surface area of fiber is small, the chloride resistance performance of FRCC with hybrid PE and steel fiber is better than that of FRCC containing only PE fiber.

Identification of seven novel virulence genes from Xanthomonas citri subsp. citri by Tn5-based random mutagenesis

Xue Song,Jing Guo,Wen-xiu Ma,Zhi-yuan Ji,Li-fang Zou,Gong-you Chen,Hua-song Zou 한국미생물학회 2015 The journal of microbiology Vol.53 No.5

To identify novel virulence genes, a mutant library of Xanthomonas citri subsp. citri 29-1 was produced using EZ-Tn5 transposon and the mutants were inoculated into susceptible grapefruit. Forty mutants with altered virulence phenotypes were identified. Nine of the mutants showed a complete loss of citrus canker induction, and the other 31 mutants resulted in attenuated canker symptoms. Southern blot analysis revealed that each of the mutants carried a single copy of Tn5. The flanking sequence was identified by plasmid rescue and 18 different ORFs were identified in the genome sequence. Of these 18 ORFs, seven had not been previously associated with the virulence of X. citri subsp. citri and were therefore confirmed by complementation analysis. Real-time PCR analysis showed that the seven genes were upregulated when the bacteria were grown in citrus plants, suggesting that the expression of these genes was essential for canker development.

Establishing a Nomogram for Stage IA-IIB Cervical Cancer Patients after Complete Resection

Zhou, Hang,Li, Xiong,Zhang, Yuan,Jia, Yao,Hu, Ting,Yang, Ru,Huang, Ke-Cheng,Chen, Zhi-Lan,Wang, Shao-Shuai,Tang, Fang-Xu,Zhou, Jin,Chen, Yi-Le,Wu, Li,Han, Xiao-Bing,Lin, Zhong-Qiu,Lu, Xiao-Mei,Xing, H Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.9

Background: This study aimed to establish a nomogram by combining clinicopathologic factors with overall survival of stage IA-IIB cervical cancer patients after complete resection with pelvic lymphadenectomy. Materials and Methods: This nomogram was based on a retrospective study on 1,563 stage IA-IIB cervical cancer patients who underwent complete resection and lymphadenectomy from 2002 to 2008. The nomogram was constructed based on multivariate analysis using Cox proportional hazard regression. The accuracy and discriminative ability of the nomogram were measured by concordance index (C-index) and calibration curve. Results: Multivariate analysis identified lymph node metastasis (LNM), lymph-vascular space invasion (LVSI), stromal invasion, parametrial invasion, tumor diameter and histology as independent prognostic factors associated with cervical cancer survival. These factors were selected for construction of the nomogram. The C-index of the nomogram was 0.71 (95% CI, 0.65 to 0.77), and calibration of the nomogram showed good agreement between the 5-year predicted survival and the actual observation. Conclusions: We developed a nomogram predicting 5-year overall survival of surgically treated stage IA-IIB cervical cancer patients. More comprehensive information that is provided by this nomogram could provide further insight into personalized therapy selection.