7^th Asia-Pacific Biotech Congress July 13-15, 2015 Beijing. China

A-Min Huang has completed his PhD from National Defense Medical Center, Taipei and Postdoctoral studies from Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. She is now an Associate Professor in the Department of Physiology, College of Medicine; National Cheng Kung University, Taiwan. She has published more than 20 papers in reputed journals in the neuroscience field.

Mechanical properties of extracellular environment are important in differentiation of neuronal stem cells and maturation of neurons. However, which genes are involved in mechanobiology of neurite outgrowth remains largely unknown. Family with sequence similarity 134 member C (Fam134c) and chromosome 3 open reading frame 10 (C3orf10) are two novel genes differentially regulate axonal and dendritic growth in rat hippocampal neurons. Fam134c is predicted as a membrane protein and C3orf10 is proposed to involve actin nucleation. Focal adhesion kinase (FAK) is a key molecule in mechanosensitivity. We hypothesized that Fam134cand C3orf10 involved in mechanobiology of neurite outgrowth under different rigidity of matrix and are up or downstream signaling molecules of FAK. Results showed that primaryrat hippocampal neurons exhibit the rigidity-dependent increases in the length of axons. Knockdown of Fam134c decreases the length of axons on glass dish but increases the length of axons on soft gel. In contrast, knockdown of C3orf10 increases the length of primary dendrites on rigid matrix. Immunointensity of both Fam134c and C3orf10 in cell bodies showed rigidity-dependent decrease. However, immunointensity of Fam134c in axons increases on soft gel but that of C3orf10 has no difference on different rigidity. Immunointensity of phosphorylated FAK (pFAK) exhibits rigidity-dependent increases in primary rat hippocampal neurons. Fam134c and C3orf10 colocalize with pFAK and paxillin on different rigidity of matrix. Knockdown of C3orf10 decreases the expression of pFAK on rigid matrix but not on soft matrix. These results provide findings that novel gene Fam134c and C3orf10 play differential roles in mechanosensing of neurons probably through the interaction with FAK.

The cure for Alzheimer’s disease suggested by the Cholinergic Hypothesis involves searching for candidate compounds that can act as inhibitors for Acetylcholinesterase (AChE) enzyme. Regional cerebral blood flow can be increased in patients with Alzheimer’s disease by Acetylcholinesterase inhibitors. In this regard, Tetraphenylporphinesulfonate (TPPS), 5,10,15,20-Tetrakis (4-sulfonatophenyl) porphyrinato Iron(III) Chloride (FeTPPS) and 5,10,15,20-Tetrakis (4-sulfonatophenyl) porphyrinatoIron(III) nitrosyl Chloride (FeNOTPPS) were investigated as candidate compounds for inhibition of Acteylcholinesterase of Drosophila melanogaster (DmAChE) by use of Molecular Docking. The results show that FeNOTPPS forms the most stable complex with DmAChE.

Tetrapyrrole compounds constitute highly conjugated systems that enable light absorption. Interaction of these photosensitizing molecules with enzymes in the interior of tumor cells has potential application in anti-cancer treatment by photodynamic therapy (PDT). Anti-cancer activity of tetrapyrrole Porphyrin derivatives was investigated by means of a molecular docking study. These molecules have the ability to reach the interior of a cell and initiate damage to intracellular molecular targets such as the P-type Ca2+-ATPase of sarcoplasmic reticulum (SERCA1a). Cache software was successfully employed for conducting docking studies of non-metallated (TPPS), metallated (FeTPPS) and metallated/nitrosylated (FeNOTPPS) Porphyrin derivatives withSERCA1a. The results show that FeTPPS forms the most stable complex.

Attention deficit hyperactivity disorder (ADHD) is a common and highly heritable disorder of childhood characterized by developmentally inappropriate inattentiveness, impulsivity, and hyperactivity. Genetic risk variants in several genes involved in regulation of dopamine and related neurotransmitter pathways are reported to be associated with ADHD. Some studies have suggested that adenosine A2A receptor (ADORA2A) plays an important role in different brain circuits and pathways involved in psychiatric disorders. Several lines of evidence implicate a relationship between mood disorder, anxiety, schizophrenia and ADORA2A gene polymorphisms. A recent study has reported that a polymorphism of ADORA2A gene is associated with attention-deficit hyperactivity disorder traits. In this study we investigated the association between the ADORA2A gene polymorphisms (rs3761422, rs35320474) and ADHD. Two samples of ADHD probands from Taiwan (n = 150) and the United Kingdom (n = 180) were genotyped and analyzed using within-family transmission disequilibrium test (TDT). We found evidence of increased transmission of the C allele of the rs3761422 polymorphism in Taiwanese samples (P=0.0018). No association was detected between the rs35320474 polymorphism and ADHD in either of the two populations. This study provides evidence for the possible involvement of ADORA2A in susceptibility to ADHD in Taiwanese population.

Dimitrov Zh has completed his PhD on 2007 at Bulgarian Academy of sciences, Institute of Molecular biology. He became an Associate Professor at University of Food Technology – Plovdiv, Bulgaria. He is a head of laboratory of Molecular biology at LB-Bulgaricum Plc. He has published more than 50 papers in area of molecular taxonomy and beneficial effects of lactic acid bacteria

Milk proteins are precursors of many biologically active peptides. These peptides are inactive within the sequence of the precursor proteins but can be released by enzymatic proteolysis during milk fermentation. Some lactic acid bacteria strains, developed in milk media, are capable to release bioactive peptides. In the present study, after evaluation of large number of strains, we selected two strains L. casei and L. bulgaricus with strong proteolytic activity including large spectra of various peptidase activities. The two strains are capable in release of Angiotensin Converting Enzyme (ACE) inhibitory peptides. A starter for Bulgarian white brined cheese, containing these strains, was constructed. The total ACE-inhibitory activity during ripening of cheese produced with the developed starter was determined in parallel with cheese produced with traditional long time used starter. The ACE-inhibitory effect was expressed as the peptide concentration needed to inhibit 50% of ACE activity (IC50). The developed starter led to considerable increase of the content of ACE-inhibitory peptides. Additionally, the total anti-oxidant activity of purified low-molecular weight peptides released by the developed starter was stronger than the one of the traditional cheese.

Michaela Jirku has graduated in 2011 at the University of Chemistry and Technology, Prague in the field of Clinical Biochemistry. She has studied PhD in biochemistry. She is a co author of 5 papers in reputed journals.

Transient receptor potential melastatin 1 (TRPM1) channel belongs to the superfamily of ion channels that respond to various physiological stimuli like chemosensation, termosensation and mechanosensation. TRP channels have six transmembrane domains with a pore region between the fifth and the sixth segment. Cytosolic N-/C-tails are responsible for regulation of TRPs which carry binding sites for signal molecules. TRPM1 channel is ubiquitously expressed in most eukaryotic cells and is involved in many cellular processes like transduction of sensory signals and regulation of Ca2+ and Mg2+ homeostasis. Mutations in TRPM1 gene effect on cell cycle in human skin cells. It seems that a loss of TRPM1 in human melanocytes correlates with increased aggressiveness in melanoma and the homozygous loss of TRPM1 in bipolar cells in retina could be a possible explanation for congenital stationary night blindness in humans.We studied possible interactions between phosphatidylinositol-4,5-bisphosphate (PIP2) and the N-terminus (NT) of TRPM1. Using bioinformatic approach we identified PIP2 binding site in A451-N566 region. Thisdomain contains several basic amino acids which can interact with anionic phospholipids. Alanine substitution mutagenesis screening revealed the crucial amino acids for these interactions. The equilibrium dissociation constants were estimated using surface plasmon resonance measurement. We identified PIP2-binding site and found mutations that decreased the affinity of TRPM1 NT/PIP2 interaction. Based on this results we concluded that basic residues playcrucial role in TRP channels binding to PIP2. Moreover, we have provided the structural insight to TRPM1- NT/PIP2 interaction using computer ligand docking

Catherine Dupré has completed her PhD at Pierre et Marie Curie University (UPMC) in 1993 before joining the Conservatoire national des arts et métiers up to 2013 as Professor Assistant. She is the co-author of 11 papers and 56 communications (speech or posters) in international congress. She has managed an industrial culture collection, Algobank-Cnam, in association with some technological transfers, particularly through two patents, to private companies. She has co-managed the R&D platform of the Cnam-Intechmer at Cherbourg, before joining as CNRS Research Engineer in 2013 the GEPEA laboratory UMR CNRS 6144 at Saint-Nazaire

Besides current applications in the nutraceutical and cosmetic sectors, microalgae and cyanobacteria have recently gained attentions for biofuel production, from biogas to biodiesel. To extend the scope of valuable microalgal products, highthroughput (HTP) screening methods are useful tools besides laboratory photobioreactors for tapping into on one hand the biodiversity (~350,000 species) and the other thephysiological flexibility of these photosynthetic microorganisms. Indeed, the achievement of competitive bioproduction through microalgae cultivation has to meet different types of requirements, from growth tests as a function of nutrients or light to identification of operational factors triggering over-accumulations of metabolites. Automated workstations with multimode readers allow microplate-based in situ and ex situ assays linked to short- and long-term incubations for respectively biochemical and growth tests. Filled 24 or 96-well microplates are incubated either in the dark for heterotrophic or under LED-panels for photoautotrophic or photoheterotrophic assays. The proposed MTS strategy has extended the use of these tools for semi-continuous cultures and some relevant other analysis. In situ measurements allow the indirect determination of cell population and pigments content and some ratios such as vitality index or carotenoid/chlorophyll after calibration of the signal on a Partial-Least-Square (PLS) regression basis, taking accounts for concentration/spectra relationships and the limits of detection and quantitation. End-point (batch cultures) or regular (semicontinuous cultures) ex-situ measurements are done through programmed plate-to-plate or plate to quartz microcells transfers for determination of some other parameters including residual nutrient concentrations, cell viability, infrared spectra, reactive oxygen species contents (ROS) for completing the required datasets.

Wenli Kang has completed her Bachelor’s study from China Agriculture University and obtained her Master’s degree from Beihang University. She is a PhD student of Nantes University supported by China Scholarship Council (CSC), and works in the laboratory GEPEA (GEnie des Procédés Environnement - Agroalimentaire) UMR CNRS 6144. She will finish her PhD study in September 2016.

Besides potential applications in the agriculture field as natural nitrogen fertilizer, N2-fixing cyanobacteria have recently gained some attentions for new applications linked to the potential production of biologically active molecules or small molecules such as hydrogen or ammonium for alga fuels. However, few data are available on cultivation of heterocytous biomass production under photoautotrophic conditions within culture medium deprived of combined nitrogen. The aim of this research was to investigate kinetic and yield parameters for biomass production by heterocytous cyanobacteria cultures in laboratory photobioreactors. The Anabaena variabilis strain PCC 7937 was studied using continuous cultures for analyzing the biomass productivity as a function of the dilution rate. Then we took advantage of chemostat cultures at a fixed dilution rate for determination of the relation between culture performances at steady-state and some environmental parameters. Optimal pH of 6.5 was determined for biomass production under N2. The experimental data confirmed that biomass productivity can be improved by increasing gas transfer efficiency and dissolved inorganic carbon concentration.

Hyun-Pil Lim is an Associate Professor and the clinical director in department prosthodontics at Chonnam National University School of Dentistry, Gwangju, Korea. He received his dental degree from Chonnam National University School of Dentistry and completed his prosthodontic training and doctorate from Chonnam National University School of Dentistry. He is the director of The Korean Academy of Prosthodontics (KAP) and Korean Academy of Stomatognathic Function and Occlusion. He has published more than 30 papers in reputed journals and has been serving as an Editorial Board Member of repute.

Peri implantitis which is one of etiologic factors associated with implant failure is mostly caused by bacteria, and can be prevented by inhibiting bacterial adhesion on implant surface. The aim of this study was to evaluate the response of Streptococcus mutans (S. mutans) and Porphyromonas gingivalis (P. gingivalis) via crystal violet staining assay on titanium surface modified by TiN sputter coating and plasma nitriding on titanium. The specimens were divided into 3 groups; polished titanium (control group), titanium modified by DC magnetron sputtering (group TiN-Ti), and titanium modified by plasma nitriding (group N-Ti). The surface characteristics of specimens were observed by using the nanosurface 3D optical profiler and field emission scanning electron microscope. Group TiN-Ti were showed TiN layer 1.2 μm in thickness and group N-Ti was identified as plasma nitriding with X-ray photoelectron spectroscopy. Roughness average (Ra) of all specimens values ≤0.2 μm, threshold Ra, which has no effect on bacterial adhesion. No significant differences of S. mutans adhesion between the surfaces of control, group TiN-Ti, and group N-Ti (P>0.05). In the result of the adhesion of P. gingivalis, there was no statistically significant difference between group TiN-Ti and control group, while group N-Ti had statistically significant higher absorbance level than the other groups (p<0.05). This indicates that too much thin modified surface could not influence the bacterial adhesion because of deficient of masking effect. Therefore TiN layer must be thick enough to make masking effect but also surface roughness values under “threshold Ra”, 0.2 μm.

Bisphenol A (BPA) is an endocrine disrupting micro pollutant. It can be biologically removed from wastewater by sorption and biodegradation. In this study, aerobic granular sludge was used, which enables a high concentration of active microorganisms per unit volume of the reactor and a high absorptive capacity due to the high content of extracellular polymers in its structure. The study determined how the concentration of BPA in wastewater influenced the transformations of nitrogen compounds. The study was conducted in five constantly aerated reactors with granules (GSBR) operated at an HRT of 16h. Influent to the control reactor did not contain BPA. BPA concentrations in the wastewater fed to the other four reactors were 2, 4, 6 and 12 mg/L. The overall COD and ammonium concentrations in synthetic wastewater were about 400 mg/L and 50 mg/L, respectively. In all reactors at least 200 cycles were performed. Ammonium nitrogen removal efficiency, resulting from nitrification and biomass synthesis, exceeded 99.5% regardless of the operational conditions. In both the control and the experimental reactors, partial nitrification (oxidation of ammonia nitrogen to nitrites) predominated. The efficiency of nitrification in the control reactor was 88.5±3.7%, and at all tested concentrations; BPA in the influent did not reduce nitrification efficiency. The highest nitrification efficiency (91.4±2.6%) was noted in the reactor fed with wastewater containing 2 mg BPA/L. In the control reactor, the efficiency of nitrogen removal from wastewater by de-nitrification and biomass synthesis was 15.8±8.6%. At concentrations of 2 and 4 mg/L, BPA in wastewater had no effect on nitrogen removal efficiency, but further increasing the concentration of BPA reduced nitrogen removal efficiency by a few percent. In the control reactor, 63% of total nitrogen removal was due to de-nitrification. In the reactor fed with wastewater containing 2 mg BPA/L, de-nitrification made the largest contribution to total nitrogen removal (70.3%).

Autophagy is an important catabolic program to response a variety of cellular stresses by forming a double membrane vesicle, autophagosome. Autophagy plays key roles in various cellular functions. Accordingly, dysregulation of autophagy is closely associated with diseases such as diabetes, neurodegenerative diseases, cardiomyopathy and cancer. In this sense, autophagy is emerging as an important therapeutic target for disease control. Among the autophagy machineries, PIK3C3/VPS34 complex functions as autophagy-triggering kinase to recruit the subsequent autophagy protein machineries on phagophore membrane. Accumulating evidence showing that inhibition of PIK3C3/VPS34 complex successfully inhibits autophagy makes the complex as an attractive target for developing autophagy inhibitors. However, one concern in PIK3C3/VPS34 complex is that many different PIK3C3/VPS34 complexes have distinct cellular functions. In this study, we have developed in vitro PIK3C3/VPS34 complex monitoring assay for autophagy inhibitor screening in a high-throughput assay format instead of targeting the catalytic activity of PIK3C3/VPS34 complex which shuts down all PIK3C3/VPS34 complexes. We performed in vitro reconstitution of an essential autophagy-promoting PIK3C3/VPS34 complex, Vps34-Beclin1-ATG14L complex in the microwell (96 well formats) plate and successfully monitored the complex formation in many different conditions. This PIK3C3/VPS34 complex protein assay would provide a reliable tool for the screening of autophagy-specific inhibitors

Yen-Po Chen got his Master’s degree in Institute of Biotechnology National Tsing Hua University (2011) in Taiwan. He is expertise in the biochemistry and cell biology of neurodegeneration. He is currently a researcher of Bio-engineering Center in Grape king BIO Ltd. His research focuses on the functional development of Trametes versicolor mycelium and neurodegenerative diseases research.

Trametes versicolor (Coriolus versicolor) is not only an edible mushroom but it has been used in traditional Chinese medicine as a supplement for therapeutic benefits for thousands of years. Historically, this mushroom was steeped in boiling water to gain access to its properties. Today, a molecular weight of about 100,000 polysaccharide peptides, PSK (polysaccharide-K), purified from Trametes versicolor mushroom mycelia from the CM-101 strain was suggested to be responsible for the biological activities. PSK (Krestin) prepared in Japan has demonstrated a significant anti-tumor effect against allogeneic tumors such as Sarcoma 180 when administered intraperitoneally or orally. Although the anti-tumor activity of PSK has been well documented, there is a paucity of information on the nerve regeneration and repair properties of PSK. Recently, the use of PSK has been linked with memory improvements in a mouse model of dementia. Therefore, we will discuss the potentials of PSK as nutraceutical or nutraceuticals to help in the reduction or even prevention of age-related neurodegenerative diseases.

I-Chen Li received her Master’sdegree in molecular medicine at National Cheng Kung University in Taiwan. She is currently the Research Specialist of Grape King Bio. Her research focuses on the development and commercialization of bio-fermentation of medicinal fungus as dietary supplements and the isolation of active compounds from the fungal mycelia.

Natural products have attained great importance as they represent a potential source of valuable secondary metabolites which can be used as food additives, nutraceuticals and pharmaceuticals. Hispidin is a secondary metabolite found in Phellinus linteus (Pl) that exhibits anticancer activity. In spite of its immense value, Pl has been shown to induce low yield of hispidin. To induce the biosynthesis of hispidin, nine Pl strains from different sources were screened under various extreme external environments to trigger the response of secondary metabolite synthesis. Due to the slow growth rate of natural Pl in the wild and the negative impact of over-deforestation on its growth, developed submerged liquid culture systems have emerged for Pl mass production. Submerged fermentation in a batch stirred tank bioreactor with constantly controlled culture parameters showed many potential advantages for both constant mycelia composition and bioactive metabolites that undergo a shorter incubation time with a lesser chance of contamination. Under optimal conditions, the production of hispidin was 830 μg/mL in a 100-L of bioreactor. We further investigate the anticancer effect of hispidin-enriched Phellinus linteus against several human cancer cells. Taken together, these results show that there is potential in developing Phellinus linteus enriched with hispidin as an ingredient in functional foods or medicinal products intended to prevent cancer.

Corticosteroid binding globulin (CBG) is alpha-globulin protein with corticosteroid binding affinity. CBG also binds several steroid hormones at high rates which mediates the physiological functions of the steroid hormones. In the present study, we demonstrated the expression of CBG gene from various tissues of four pig breeds including Berkshire, Duroc, Landrace, and Yorkshire. Up-regulation of CBG mRNA was detected in liver of all four breeds. Interestingly, the mRNA of CBG in liver of Berkshire showed the highest expression compared with that of other breeds. We also examined single nucleotide polymorphisms (SNPs) of CBG and the CBG SNP c.882 A>G; 265I>V was found from the Berkshire liver, and then we analyzed the association between SNP and various meat quality traits. The SNP was significantly associated with backfat thickness, pH24hr, CIE a*, CIE b*, water holding capacity, fat content, moisture content, protein content, cooking loss, and shear force. We also analyzed the association between genotype of CBG and meat quality traits depending on sex. The SNP of CBG showed significant differences on various meat quality traits including backfat thickness, pH24hr, CIE a*, CIE b*, water holding capacity, fat content, moisture content, protein content, drip loss, and cooking loss in gilt whereas the SNP was only involved in cooking loss and shear force in barrow. In conclusion, we suggest that the analysis of SNP might be a molecular maker for improvement of pork production.

Feather is a waste from the poultry processing industry and it contains over 90% crude protein and about 15% N. Feather meals have a potential use as a slow-release N fertilizer for traditional agricultural practices and organic agricultural practices. Therefore, degradation of feather to feather meal is important for agricultural applications. Steam pressure cooked method and chemical treatment method have been used to produce feather meals, however, the meals produced under these methods showed low agricultural values. Biodegradation of feather waste by microorganisms is a viable alternative. A screening program to isolate micro-organisms exhibiting keratinase activity has been conducted in this laboratory. Several bacteria isolates showed good ability to degrade feathers will be discussed in this conference

An example of an endocrine disrupting micro pollutant that can be biologically removed from wastewater by sorption and biodegradation is bisphenol A (BPA). When biologically removing BPA, granular biomass allows a high concentration of active microorganisms per unit volume of the reactor and a high absorptive capacity due to the high content of extracellular polymers in its structure. To determine how the concentration of BPA in wastewater influences the effectiveness of carbon compounds (COD) removal, biomass growth (Y) and granule diameter, five constantly aerated reactors with granules (GSBRs) were operated at an HRT of 16 h. BPA concentrations in the synthetic wastewater were 0 mg/L in the control reactor, and 2, 4, 6 and 12 mg/L in the remaining four reactors. The overall COD concentration in the influent to the reactors, including BPA concentration, was about 400 mg COD/L; ammonium concentration was about 50 mg N-NH4/L. At least 200 cycles were performed in all reactors. Biomass concentration in the GSBRs ranged from 9268±2266 mg TSS/L at a BPA concentration of 2 mg/L to 10448±2266 mg TSS/L at a BPA concentration of 12 mg/L. The diameter of the granules increased with increasing BPA concentration in the influent. At the two highest BPA concentrations, granules with the diameter over 2 mm dominated in the biomass. These granules accounted for 24% and 37% of the total biomass, respectively at the BPA concentrations of 6 mg/L and 12 mg/L in the influent. This greater size may have allowed for better retention of biomass, thus slightly increasing the biomass concentration. Regardless of BPA concentrations in wastewater, COD was removed within the first 1-1.5 h of the GSBRs cycle. The effectiveness of COD removal by the aerobic granular sludge ranged from 75.1±17.8% at a concentration of 4mg BPA/L to 91.4±7.7% at a concentration 12 mg BPA/L, which may indicate that the latter concentration of BPA in the influent increased the effectiveness of COD removal. Y was lowest (0.28g TSS/g COD) at concentrations of 2 and 12 mg BPA/L and highest (0.42g TSS/g COD) at 6 mg BPA/L, as compared to 0.38g TS/g COD in the control reactor.

Bisphenol A (BPA), an endocrine disrupting compound, is almost completely removed from wastewater through biological processes in activated sludge or immobilized biomass. However, the biological effluent may contain high concentrations of suspended solids or organic compounds (COD). In this study, wastewater treated in granular sequencing batch reactors (GSBRs) was characterised by COD concentrations from 107.1 to 145.5 mg/L and suspended solid concentrations from 101 to 171 mg/L. This wastewater (feed solution) was post-treated with ceramic membranes that are characterised by mechanical and chemical stability, long life and good antifouling properties. Microfiltration (MF) and ultrafiltration (UF) were conducted at transmembrane pressures (TMP) of 0.2 MPa and 0.3 MPa, respectively. Both MF and UF caused complete rejection of suspended solids. COD rejection during MF was 81.1±1.1%, independently of the composition of the feed solution. The use of UF did not increased COD rejection significantly. During MF, from 34 to 60% of COD was adsorbed to the membrane surface, as determined from the mass balance of membrane filtration. The use of UF increased COD adsorption to 71%. Permeate flux changed from 24.0 to 29.5 L/(m2∙h), and volumetric concentration factor ranged from 2.0 to 2.4; these hydraulic parameters were influenced neither by the type of filtration process nor by TMP. Although filtration capacity decreased with time because of fouling, this decrease was not affected by the composition of the feed solution. These results suggest that the use of both MF and UF ceramic membranes is a good solution for post-treatment of secondary effluent from GSBRs that treat wastewater containing BPA. However, taking everything into consideration, MF is more advantageous because it can be operated at lower TMP, thus requiring less energy for filtration.