Repeated-Batch and Continuous Production of L-Lactic Acid by Rhizopus oryzae Immobilized in Calcium Alginat Beads:ReactorPerformance and Kinetic Model

Repeated-batch and continuous production of L-lactic acid by immobilized Rhizopusoryzae with calcium alginate entrapment method in a three-phase fluidized-bed bioreactor was stud-ied.The operation conditions were optimized.The productivity based on total reactor volume wasabout 3 times higher than that with free cells in a traditional stirred tank bioreactor.A mathemat-ical model was proposed and the model predictions were in good agreement with the experimentaldat.

Synthesis of the Biomimetic Polymer: Aliphatic Diamine and RGDS Modified Poly(d,l-lactic acid)

A novel poly（d, /-lactic acid） （PDLLA） based biomimetic polymer was synthesized by grafting maleic anhydride, butanediamine and arg-gly-asp-ser （RGDS） peptides onto the backbone of PDLLA, aiming to overcome the acidity and auto-accelerating degradation of PDLLA during degradation and to improve its biospecificity and biocompatibility. The synthetic copolymer was characterized by FTIR, ^13C NMR and amino acid analyzer （AAA）.

Optimization of L-lactic Acid Production of Rhizopus Oryzae Mutant RLC41-6 by Ion Beam Implantation at Low-Energy

In order to obtain an industrial strain with a higher L（＋）-lactic acid yield, the strain Rhizopus oryzae RF3608 was mutated by means of nitrogen ion beam implantation and the mutant strain RLC41-6 was isolated. Under optimal conditions the yield of L（＋）-lactic acid produced in a shake-flask reached 133 g/L - 137 g/L after 36 h cultivation, indicating that the conversion rate based on glucose was as high as 88% - 91% and the productivity was 3.75 g/L.h. It was almost a 115% increase in lactic acid production compared with the original strain RF3608.

Optimization of L(+)-Lactic Acid Production from Xylose with Rhizopus Oryzae Mutant RLC41-6 Breeding by Low-Energy Ion Implantation

In order to obtain an industrial strain with a higher L（＋）-lactic acid yield, the strain Rhizopus oryzae PW352 was mutated by means of nitrogen ion beam implantation and the mutant strain Rhizopus oryzae RLC41-6 was obtained. An experimental finding was made in surprise that Rhizopus oryzae mutant RLC41-6 is not only an L（＋）-lactic acid producer from corn starch but also an efficient producer of L（＋）-lactic acid from xylose. Under optimal conditions, the production of L（＋）-lactic acid from 100 g/L xylose reached 77.39 g/L after 144 h fed-batch fermentation, A high mutation rate and a wide mutation spectrum of low-energy ion implantation were observed in the experiment.

Improvement of L(+)-Lactic Acid Production of Rhizopus Oryzae by Low-Energy Ions and Analysis of Its Mechanism

The wild type strain Rhizopus oryzae PW352 was mutated by means of nitrogen ion implantation （15 keV, 7.8×10^14 ～ 2.08 ×10^15 ions/cm^2） to find an industrial strain with a higher L（＋）-lactic acid yield, and two mutants RE3303 and RF9052 were isolated. In order to discuss the mechanism primarily, Lactate Dehydrogenase of Rhizopus oryzae was studied. While the two mutants produced L（＋）-lactic acid by 75% more than the wild strain did, their specific activity of Lactate Dehydrogenase was found to be higher than that in the wild strain. The optimum temperature of Lactate Dehydrogenase in Rhizopus oryzae RF9052 was higher. Compared to the wild strain, the Michaelis constant （Km） value of Lactate Dehydrogenase in the mutants was Changed. All these changes show that L（＋）-lactic acid production has a correlation with the specific activity of Lactate Dehydrogenase. The low-energy ions, implanted into the strain, may improve the specific activity of Lactate Dehydrogenase by influencing its gene structure and protein structure.

L-LACTIC ACID FERMENTATION IN A ROTATING-DISC CONTACTOR WITH SIMULTANEOUS PRODUCT SEPARATION BY ION-EXCHANGE

A rotating disk contactor（RDC）was designed to perform L-lactic acid fermentation with afilamentous fungi,Rhizopus oryzae,which was immobilized on the surfaces of the rotating discs.Thebioreactor was operated using repeated-batch method as well as continuous feeding method.Ananionic resin,D354,slightly basic in nature and of high selectivity and capacity was chosen for lacticacid separation.A coupled process of L-lactic acid fermentation and ion-exchange separation wasevaluated experimentally.The results indicated that the pH value of the fermentation broth could bemaintained at about 3-3.5 without any addition of alkali.The conversion ratio of glucose to L-lacticacid was about 0.7 g·g-1 and the fermentation rate reached as high as 62.5 g·h-1·m-2.

Synthesis and Characterization of Chitosan-g-poly- (D, L-lactic acid) Copolymer

Biodegradable chitosan-g-poly (D, L-lactic acid) copolymers were prepared via two methods. (1) The lactide was grafted onto hydroxyl groups of chitosan by using macromolecular initiator sodium of trimethylsilyl-chitosan, (2) poly (D,L-lactic acid)(PLA) with low molecular weight can be linked to the amino group by coupling activated PLA to trimethylsilyl-chitosan. Two graft copolymers had hydrophilic-hydrophobic character and can be applied as carriers for drug delivery.

Enhancement of L(+)-Lactic Acid Production of Immobilized Rhizoous Orvzae Imolanted by Ion Beams

Immobilized Rhizopus oryzae culturing may be a solution to the inhibited production of L（＋）-lactic acid in submerged fermentation, which is caused by aggregated mycelia floc. In the present study, a R. oryzae mutant （RL6041） with a 90% conversion rate of glucose into L-lactic acid was obtained by N＋ implantation under the optimized conditions of a beam energy of 15 keV and a dose of 2.6 ×10^15 ions/cm^2. Using polyurethane foam as the immobilization matrix, the optimal L-lactic acid production conditions were determined as 4 mm polyurethane foam, 150 r/min, 50 g/L ～ 80 g/L of initial glucose, 38℃ and pH 6.0. 15-cycle repeated productions of L-lactic acid by immobilized RL6041 were performed under the optimized culturing conditions and over 80% of the glucose was converted into L-lactic acid in 30 hours on average. The results show that immobilized RL6041 is a promising candidate for continuous L-lactic acid production.

SURFACE OF GELATIN MODIFIED POLY(L-LACTIC ACID)FILM

In this paper, the surface structure of poly(L-lactic acid) (PLLA) film modified with gelatin was investigated. ThePLLA film specimens were treated directly with aqueous alkali solution to provide their surfaces with carboxyl groups, sothat these functional groups could become the reactive sites for gelatin immobilization. The functional groups of the PLLAfilms were identified by ATR-FTIR spectra and XPS spectra, the changes in surface morphology were observed by usingenvironmental scanning electron microscopy (ESEM), and the hydrophilicity of modified PLLA films was examined bywater contact angle measurement. Experimental results showed that the gelatin was immobilized with water-solublecarbodiimide (EDC) onto the PLLA film's surfaces, and the gelatin content on the polymer surface was related to carboxylicgroup formed in the controlled hydrolysis process. Rough surfaces caused by hydrolysis will predominantly favor the adhesion and growth of cell; and the hydrophilicity of these surfaces after the modification procedure is enhanced.

Sorption and Diffusion Behavior of Carbon Dioxide into Poly（l-lactic acid） Films at Elevated Pressures

Equilibrium sorption amount, desorption diffusion coefficients and sorption diffusion coefficients of CO2 in poly（l-lactic acid） （PLLA） films at elevated pressures were determined by the gravimetric method, in which the Fick＇s diffusion model was applied to analyze both the desorption and sorption processes. The equilibrium sorption amount of CO2 in PLLA increased with lowering temperature and elevating pressure at the temperature range from 40 to 60 ℃ and pressure from 10^4 to 2x10^4 kPa. Desorption diffusion coefficients were greatly influenced by the equilibrium sorption amount, and they were in the same order of magnitude as the sorption diffusion coefficients. The scan electron microscope （SEM） photos demonstrated that there was no foaming phenomenon of the PLLA film during desorption and sorption processes. The XRD spectra implied that the crystalline degree of PLLA film decreased after CO2 processing. It was concluded that PLLA polymer could be well swollen and plasticized by supercritical CO2.

Optimization of L(+)-Lactic Acid Fermentation Without Neutralisation of Rhizopus Oryzae Mutant RK02 by Low-Energy Ion Implantation

In order to get an industrial strain which can yield a high concentration of lactic acid for ISPR （in situ product removal）, the original strain Rhizopus oryzae RE3303 was mutated by low-energy ion beam implantation. A mutant RK02 was screened, and the factors such as the substrate concentration, nitrogen source concentration, inoculum size, seed age, aeration and temperature that affect the production of lactic acid were studied in detail. Under optimal con- ditions, the maximum concentration of L（＋）-lactic acid reached 34.85 g/L after 30 h shake-flask cultivation without adding any neutralisation （5% Glucose added）, which was a 146% increase in lactic acid production after ion implantation compared with the original strain. It was also shown that RK02 can be used in ISPR to reduce the number of times of separation.

Chitosan——L-Lactic Acid Scaffold for the Regeneration of Peripheral Nerve and Its NGF Release Properties

Chitosan—L-lactic acid composite scaffold for the regeneration of peripheral nerve is obtained by grafting L-lactic acid onto the amino groups in chitosan with combined vacuum freezer drier. The composite scaffold was characterized by ATR-FTIR and SEM. The scaffold has a better graft efficiency and has a dense inner layer and a loose outer layer with porous structure, and the pore size is about 100 μm.The NGF release properties of the scaffold were investigated. The experimental results showed that, at the 1st day, 15.2 ng of NGF on average was released from the scaffold. From day 2 to day 10, the release rate obviously slowed down and 1.64 ng of NGF was released on average every day. After 10 days, the release rate was slower and 10.3 ng of NGF was released on average every day. After 60 days, NGF could also maintained a certain concentration. These properties show that the scaffold is a better carrier for NGF which can be more advantageous to the regeneration of the damaged peripheral nerve. As a result, this composite scaffold would be an ideal candidate for the regeneration of damaged peripheral nerve.

Multi-porous electroactive poly(L-lactic acid)/ polypyrrole composite micro/nano fibrous scaffolds promote neurite outgrowth in PC12 cells

In this study, poly（L-lactic acid）/ammonium persulfate doped-polypyrrole composite fibrous scaffolds with moderate conductivity were produced by combining electrospinning with in situ polymerization. PC12 cells were cultured on these fibrous scaffolds and their growth following electrical stimulation （0-20.0 μA stimulus intensity, for 1-4 days） was observed using inverted light microscopy, and scanning electron microscopy coupled with the MTT cell viability test. The results demonstrated that the poly（L-lactic acid）/ammonium persulfate doped-polypyrrole fibrous scaffold was a dual multi-porous micro/nano fibrous scaffold. An electrical stimulation with a current intensity 5.0- 10.0 μAfor about 2 days enhanced neuronal growth and neurite outgrowth, while a high current intensity （over 15.0 μA） suppressed them. These results indicate that electrical stimulation with a moderate current intensity for an optimum time frame can promote neuronal growth and neurite outgrowth in an intensity- and time-dependent manner.

Preparation of Poly ( L-lactic acid ) / Vermiculites Composites by in situ Copolymerization of Lactic Acid and Hydroxylic Vermiculites

L-lactic acid (L-LA) based copolymer/hydroxylation vermiculites composites (PLLA-co-bis A/HVMTs) were prepared by in situ reaction among L-LA, adipic acid, and hydroxylation lamellar vermiculites (HVMTs) using bisphenol-A epoxy resin as chain extending agent. HVMTs were obtained by sulfuric acid-leaching of lamellar vermiculites (VMTs). The effects of sulfuric acid leaching on the VMTs structure were characterized by X-ray diffraction (XRD), 29Si magic-angle spinning nuclear magnetic resonance(29Si NMR), Fourier transform infrared spectroscopy (FTIR), field emission scanning electron microscopy (FE-SEM), and thermogravimetric analysis (TGA). FTIR, FE-SEM, and TGA were used to characterize the reaction activity of HVMTs. The results indicated that VMTs with increased hydroxyl groups had been successfully obtained and could react with -COOH of the reaction system. The amount of L-LA based copolymer grafted on the surface of HVMTs was more than 22%. The onset decomposition temperature of L-LA based copolymer grafted on the surface of HVMTs was 30℃ higher than that of free L-LA based copolymer.

Effects of cultivation conditions on the optical purity of L(+)-lactic acid produced from corncob hydrolysate by Rhizopus oryzae

The effect ofcultivation conditionson the opticalpurity ofL(+)-lactic acid produced by Rhizopusoryzae HZS6 from corncob hydrolysate was investigated. The isomeric composition of lactic acid was influenced by the supplementation of L(+)-lactic acid to fermentation medium. L(+)-Isomer increased with the dosage,no(-)-lactic acid was observed when the concentration of supplemented L(+)-lactic acid in matrix was≥1.5g l-1. However,the L(+)-lactic acid yield decreased with the dosage. Under suitable conditions,100g l-1 initial corncob xylose,2g l-1 NH4NO3,1.5 g l-1 supplemented L(+)-lactic acid,R. oryzae HZS6 could produce 100% L(+)-form lactic acid with the yield of 75% and final concentration of 81.2 g l-1,at pH 6.0 and temperature 34℃.

Transplantation of Nogo-66 receptor gene-silenced cells in a poly(D,L-lactic-co-glycolic acid) scaffold for the treatment of spinal cord injury

Inhibition of neurite growth, which is in large part mediated by the Nogo-66 receptor, affects neural regeneration following bone marrow mesenchymal stem cell transplantation. The tissue engineering scaffold poly（D,L-lactide-co-glycolic acid） has good histocompatibility and can promote the growth of regenerating nerve fibers. The present study used small interfering RNA to silence Nogo-66 receptor gene expression in bone marrow mesenchymal stem cells and Schwann cells, which were subsequently transplanted with poly（D,L-lactide-co-glycolic acid） into the spinal cord lesion regions in rats. Simultaneously, rats treated with scaffold only were taken as the control group. Hematoxylin-eosin staining and immunohistochemistry revealed that at 4 weeks after transplantation, rats had good motor function of the hind limb after treatment with Nogo-66 receptor gene-silenced ceils prus the poly（O,L-lactide-co-glycolic acid） scaffold compared with rats treated with scaffold only, and the number of bone marrow mesenchymal stem cells and neuron-like cells was also increased. At 8 weeks after transplantation, horseradish peroxidase tracing and transmission electron microscopy showed a large number of unmyelinated and myelinated nerve fibers, as well as intact regenerating axonal myelin sheath following spinal cord hemisection injury. These experimental findings indicate that transplantation of Nogo-66 receptor gene-silenced bone marrow mesenchymal stem cells and Schwann cells plus a poly（D,L-lactide-co-glycolic acid） scaffold can significantly enhance axonal regeneration of spinal cord neurons and improve motor function of the extremities in rats following spinal cord injury.

Callus Induction of Alfalfa and Evaluation of Callus Tolerance to Acid and Aluminum

Medicago is an important grass in the legume family,which is suitable to be grown in neutral or alkalescent soils,and hence,can be widely spread in south of China with low pH value.It is one of major objectives to cultivate acid-aluminum tolerant species.In the research,6 high-quality species with high-yielding potential were taken as materials to make evaluation on acid-aluminum tolerance by cell-culture technique,and the results showed that tolerance capacity from high to low was GT13R＞Muge 701=Muge 702＞Acrora＞AC-3＞Sheshou No.2=Medicago sativa.

Variations in Phytic Acid Content and Their Relationship with Protein Content and Kernel Morphological Characters of Chinese Winter Wheat Cultivars

[Objective] The aim of this study was to determine the variation of phytic acid content （PAC） and its frequency distribution, as well as the relationship between PAC and protein content, kernel characteristics in 161 Chinese winter wheat cultivars from four regions. [Method] One hundred and sixty-one winter wheat varieties from China Wheat Zones I, II, III and IV （Table 1） were grown in a randomized block de- sign, in the 2009-2011 cropping season; and then the indexes for describing the grain morphological characteristics such as the thousand kernel weight （TKW）, kernel length （KL）, kernel width （KW） and kernel thickness （KT） were measured; the phytic acid content （PAC）, protein content and sedimentation value were also determined; finally, the relationship between PAC and protein content, kernel characteristics were analyzed. [Result] The PAC in the cultivars tested ranged from 0.92% to 1.95% with a mean value of 1.41%. Protein content ranged from 12.60% to 19.20%, with a mean of 15.24%. Most （53.4%） of the wheat genotypes had a PAC value in the range of 1.25% to 1.55%. No significant correlation was found between PAC and protein content, sedimentation value, while protein content and SDS sedimentation value was significant correlated, which suggested the possibility of breeding wheat cultivars that have a low PAC but a high protein content and good gluten quality. There was a high correlation between TKW and KW （,.=0.79）, KL （r=0.50） and KT （r=0.64）. PAC was found having no significant correlation with TKW, KW, KL and KT. [Conclusion] The result suggests the possibility of breeding wheat cultivars that have a low PAC but high kernel weight.

Corrosion properties of stainless steel 316L/Ni-Cu-P coatings in warm acidic solution

In order to improve corrosion resistance of stainless steel 316L in warm acidic solution, Ni？Cu？P coatings with high copper and phosphorus contents were deposited onto stainless steel 316L substrates via electroless plating. The structure of the film and its resistance to corrosion in a warm acidic environment were investigated using scanning electron microscopy （SEM）, energy dispersive spectroscopy （EDS）, X-ray diffraction spectrometry （XRD）, polarization curves, electrochemical impedance spectroscopy （EIS）, and dipping corrosion tests, respectively. The results demonstrate that Ni？Cu？P coatings consist of two types of nodules, which are 19.98% Cu and 39.17% Cu （mass fraction） respectively. The corrosion resistance of the 316L substrate when subjected to a warm acidic solution is significantly improved by the addition of the new type of the Ni？Cu？P coating. The as-plated coatings demonstrate better corrosion resistance than annealed coatings. As-plated coatings and those annealed at 673 K are found to corrode selectively, while pitting is observed to be the main corrosion mechanism of coatings annealed at 773 and 873 K.

HPLC法同时测定杠板归不同部位中5种成分的含量

目的建立高效液相色谱法同时测定杠板归不同部位(茎、叶、根)中5种成分的含量。方法采用Luna?-C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸水为流动相梯度洗脱,流速为1.0 mL/min,检测波长为320 nm,柱温为30℃,测定杠板归不同部位中绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素的含量。结果绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素分别在0.046~4.6μg/mL,0.212~21.2μg/mL,0.055~5.5μg/mL,0.084~8.4μg/mL,0.114~11.4μg/mL范围内质量浓度与峰面积呈线性关系良好(r>0.9990),平均加样回收率为97.65%~102.09%,RSD为1.76%~2.44%。不同部位中5种成分含量差异较大,绿原酸和咖啡酸2个成分在叶中含量高,且远远高于根和茎中的含量;芦丁和阿魏酸主要存在于茎和根中,叶中未检测到;槲皮素在茎中含量最高,叶中次之,根中最低。结论建立的HPLC方法准确可靠、重复性好、专属性强,可用于杠板归不同部位中5个成分含量的同时测定。