Peilin Cen

Butyric acid fermentation in a fibrous bed bioreactor with immobilized Clostridium tyrobutyricum from cane molasses

Butyrate fermentation by immobilized Clostridium tyrobutyricum was successfully carried out in a fibrous bed bioreactor using cane molasses. Batch fermentations were conducted to investigate the influence of pH on the metabolism of the strain, and the results showed that the fermentation gave a highest butyrate production of 26.2 g l(-1) with yield of 0.47 g g(-1) and reactor productivity up to 4.13 g l(-1)h(-1) at pH 6.0. When repeated-batch fermentation was carried out, long-term operation with high butyrate yield, volumetric productivity was achieved. Several cane molasses pretreatment techniques were investigated, and it was found that sulfuric acid treatment gave better results regarding butyrate concentration (34.6+/-0.8 g l(-1)), yield (0.58+/-0.01 g g(-1)), and sugar utilization (90.8+/-0.9%). Also, fed-batch fermentation from cane molasses pretreated with sulfuric acid was performed to further increase the concentration of butyrate up to 55.2 g l(-1).

Recent advances in the research and development of human defensins

Human defensins are a family of cationic antimicrobial peptides with molecular weights of 4-5 kDa, containing a conserved six disulphide-linked cysteine motif. During the last two decades a series of endogenous alpha- and beta-human defensins were discovered. They exhibit a broad range of antimicrobial properties and are thought to be ideal therapeutic agents because of their potential ability to circumvent the problems of acquired resistance often observed with other antimicrobial therapies. Because of their appealing medical and pharmaceutical potential there has been an emphasis on human defensins in medical and molecular biology research in recent years. This paper aims to present a comprehensive review of recent advances in the study of human defensins including their discovery, classification, molecular properties, expression, mechanisms of action and potential medical applications. In addition, the advances in producing human defensins via genetic engineered cells are summarized from research works in our group (besides host cells including E. coli, B. subtilis and yeast systems, the cell-free protein synthesis system was also employed to express human beta-defensin-2) along with other related published works. The present challenges and prospects for the potential application of human defensins are also discussed.

Kinetics of Levulinic Acid Formation from Glucose Decomposition at High Temperature

Abstract Levulinic acid is a kind of new green platform chemical with wide application. The kinetics of levulinic acid formation from glucose decomposition at high temperature was investigated. Glucose containing 1%, 3% or 5% H 2 SO 4 was treated at 170°C or 190°C. For the various experimental conditions assayed, the time-courses of glucose and glucose degradation products (including 5-hydroxymethylfurfural and levulinic acid) were established. These variables were correlated with the reaction time based on the equations derived from a pseudo-homogeneous, first-order kinetic model, which provided a satisfactory interpretation of the experimental results. The set of kinetic parameters from regression of experimental data provided useful information for understanding the levulinic acid formation mechanism.

Enhanced butyric acid tolerance and bioproduction by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor

Repeated fed‐batch fermentation of glucose by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor (FBB) was successfully employed to produce butyric acid at a high final concentration as well as to adapt a butyric‐acid‐tolerant strain. At the end of the eighth fed‐batch fermentation, the butyric acid concentration reached 86.9 ± 2.17 g/L, which to our knowledge is the highest butyric acid concentration ever produced in the traditional fermentation process. To understand the mechanism and factors contributing to the improved butyric acid production and enhanced acid tolerance, adapted strains were harvested from the FBB and characterized for their physiological properties, including specific growth rate, acid‐forming enzymes, intracellular pH, membrane‐bound ATPase and cell morphology. Compared with the original culture used to seed the bioreactor, the adapted culture showed significantly reduced inhibition effects of butyric acid on specific growth rate, cellular activities of butyric‐acid‐forming enzyme phosphotransbutyrylase (PTB) and ATPase, together with elevated intracellular pH, and elongated rod morphology. Biotechnol. Bioeng. 2011; 108:31–40.

Advances in the Research and Development of Acrylic Acid Production from Biomass

Abstract The shortage of petroleum has resulted in worldwide efforts to produce chemicals from renewable resources. Among these attempts, the possibility of producing acrylic acid from biomass has caught the eye of many researchers. Converting the carbohydrates first to lactic acid by fermentation and then dehydrating lactic acid to acrylic acid is hitherto the most effective way for producing acrylic acid from biomass. While the lactic acid fermentation has been commercialized since longer times, the dehydration process of lactic acid is still under development because of its low yield. Further efforts should be made before this process became economically feasible. Because of the existence of acrylic acid pathways in some microorganisms, strain improvement and metabolic engineering provides also a possibility to produce acrylic acid directly from biomass by fermentation.

Production of Cellulase by Solid-State Fermentation

The advances in the production of cellulase by solid-state fermentation are reviewed. In the process description of solid-state fermentation for cellulase production, the microorganisms, raw materials for solid-state fermentation, pretreatment of raw materials, sterilization and inoculation, and solid-state fermentation process are included. The effects and control of operating conditions, such as temperature, water content and water activity, pH, aeration, and substrate, are presented. A total of eight types of bioreactors with their advantages and disadvantages for solid-state fermentation are discussed. From the engineering aspect, mathematical models for the solid-state fermentation process are also discussed and several suggestions are proposed which might further research and development of cellulase production by the solid-state fermentation process.

Kinetics of Simultaneous Saccharification and Lactic Acid Fermentation Processes

After pretreatment of corn cob by dilute acid, the lignocellulosic residue was used as raw materials for the simultaneous saccharification and lactic acid fermentation (SSLF). Because of the same optimal temperature and pH requirement as well as the anaerobic condition, the lactic acid fermentation is perfectly compatible with enzymatic hydrolysis of cellulosic materials. In the SSLF processes, the final concentration of lactic acid reached 33.97g/L with a conversion ratio of 79% based on the consumed cellulose. A mathematical model is suggested to simulate the SSLF process with good agreement.

A high-throughput method for screening of rapamycin-producing strains of Streptomyces hygroscopicus by cultivation in 96-well microtiter plates.

A novel high-throughput cultivation method was developed to rapidly screen large numbers of rapamycin-producing mutants of Streptomyces hygroscopicus by duplicate culturing of isolates on the surfaces of agar-solidified 96 wells in microtiter plates. One copy of the cultures was used for the rapamycin bioassay and the other identical copy, representing potentially high yielding strains, was preserved for further study. By integrating 96-well solid cultivation and the bioassay, we screened more than 7000 isolates and found 10 high-yielding strains. From these, one mutant produced 420 μg rapamycin/ml, which was double the yield of parent strain used in the submerged fermentation process.

Kinetic Studies on Wheat Straw Hydrolysis to Levulinic Acid

Abstract Levulinic acid is considered as a promising green platform chemical derived from biomass. The kinetics of levulinic acid accumulation in the hydrolysis process of wheat straw was investigated in the study. Using dilute sulfuric acid as a catalyst, the kinetic experiments were performed in a temperature range of 190–230°C and an acid concentration range of 1%–5% (by mass). A simple model of first-order series reactions was developed, which provided a satisfactory interpretation of the experimental results. The kinetics of main intermediates including sugar and 5-hydroxymethylfurfural (5-HMF) were also established. The kinetic parameters provided useful information for understanding the hydrolysis process.

Preparative Scale Production of Functional Mouse Aquaporin 4 Using Different Cell-Free Expression Modes

The continuous progress in the structural and functional characterization of aquaporins increasingly attracts attention to study their roles in certain mammalian diseases. Although several structures of aquaporins have already been solved by crystallization, the challenge of producing sufficient amounts of functional proteins still remains. CF (cell free) expression has emerged in recent times as a promising alternative option in order to synthesize large quantities of membrane proteins, and the focus of this report was to evaluate the potential of this technique for the production of eukaryotic aquaporins. We have selected the mouse aquaporin 4 as a representative of mammalian aquaporins. The protein was synthesized in an E. coli extract based cell-free system with two different expression modes, and the efficiencies of two modes were compared. In both, the P-CF (cell-free membrane protein expression as precipitate) mode generating initial aquaporin precipitates as well as in the D-CF (cell-free membrane protein expression in presence of detergent) mode, generating directly detergent solubilized samples, we were able to obtain mg amounts of protein per ml of cell-free reaction. Purified aquaporin samples solubilized in different detergents were reconstituted into liposomes, and analyzed for the water channel activity. The calculated P f value of proteoliposome samples isolated from the D-CF mode was 133 µm/s at 10°C, which was 5 times higher as that of the control. A reversible inhibitory effect of mercury chloride was observed, which is consistent with previous observations of in vitro reconstituted aquaporin 4. In this study, a fast and convenient protocol was established for functional expression of aquaporins, which could serve as basis for further applications such as water filtration.