Production of acid proteinase by Humicola lutea 120-5 immobilized in mixed photo-cross-linked polyvinyl alcohol and calcium-alginate beads

Humicola lutea 120-5 spores were immobilized in a mixed photo-crosslinked polyvinyl alcohol and calcium-alginate gel. Maximum enzyme synthesis was established with 1:8 (v:v) gel beads: growth medium inoculum and 48 h duration of one cycle. The free cells were very unstable in replacement fermentations. The operational stability of the immobilized system indicated the possibility of the application of Humicola lutea 120-5 in a semi-continuous process for the production of acid proteinase.     

Reject water treatment by improvement of whole cell anammox entrapment using polyvinyl alcohol/alginate gel

Reject water treatment performance was investigated by whole cell anammox sludge entrapped polyvinyl alcohol/sodium alginate gel in the stirred tank reactor (STR). The whole experiment was conducted through Phase 1 and Phase 2 in which synthetic wastewater and modified reject water were used as feeding medium, respectively. The anammox reactor demonstrated quick start-up after 22 days as well as stable and relatively high nitrogen removal rate of more than 8.0 kg-N m⁻³ day⁻¹ during the two both phases even under moderately low temperature of 25 ± 0.5°C during the last 2 months of Phase 2. The matured brownish red PVA beads had good characteristics with buoyant density of 1.10 g cm⁻³, settling velocity of 141 m h⁻¹ and diameter of 4 mm. The bacterial community was identified by 16S rDNA analysis revealing the concurrent existence of KSU-1 and new kind anammox bacterium Kumadai-I after changing influent from synthetic wastewater to reject water. It was speculated that Kumadai-I might play a role as “promotion” factor together with KSU-1 on high nitrogen removal rate. These results demonstrate the potential application of whole cell anammox entrapment by PVA/alginate gel for achieving stable and high-rate nitrogen removal from high ammonium with low C/N ratio contained wastewaters, such as reject water, digester liquor or landfill leachate.     

Evidence for egg-box-compatible interactions in calcium-alginate gels from fiber X-ray diffraction

The structures of guluronic-acid-rich alginate in the acid and calcium forms were investigated using fiber X-ray diffraction. Data recorded for alginate fibers in the acid form show a repeat along the chain axis of c = 0.87 nm, a value that is in agreement with the one measured by Atkins et al. (Biopolymers 1973, 12, 1865) and contradicts a repeat of 0.78 nm recently suggested by Li et al. (Biomacromolecules 2007, 8, 464). In the Ca2+ form, our observations indicate that the junction zone involves dimerization of polymer chains through Ca2+ coordination according to the egg-box model. For reasons that are not understood at present, coordination of the divalent cations reduces the ability for the lateral crystallographic packing of the dimers. A proposed model for the junction zone involves polymer chains packed on a hexagonal lattice with a lattice constant a = 0.66 nm. Random pairs of chains form dimers through coordination of Ca2+ cations. Further lateral interaction between dimers is mediated by disordered Na+ and Ca2+ cations, water molecules, and hydrogen bonding.     

Immobilization of microbial cells using polyvinyl alcohol (PVA) — polyacrylamide gels

Summary A novel cell immobilization technique is developed in which polyvinyl alcohol is crosslinked with boric acid with addition of an acrylamide as a polymerizing agent. The presence of the polymerizing agent overcomes the problem of swelling of PVA gels in aqueous solution. The new immobilization method was used to entrap a phenol-degrading microorganism, a species of Pseudomonas. Phenol was successfully degraded in a shake-flask test of the beads demonstrating that cell viability was maintained following the immobilization procedure.     

聚乙烯醇脱氢酶的提取及检测

为解决结合在细胞上的可溶性蛋白聚乙烯醇脱氢酶（PVADH）的检测困难问题,从提取及检测两方面对该酶进行研究,并对检测方法进行改进。结果表明,非离子型表面活性剂Triton X-100对可溶性蛋白PVADH的提取效果优于离子型表面活性剂炕基苯磺酸钠（LAS）和溴化十六烷基吡啶（CPB）,酶活力比LAS和CPB提取后所得酶活力分别提高246．5％和831．3％。而非离子型表面活性剂中,Triton X-100与Tween80相比,所得最高酶活提高了101．4％。Triton X—100浓度和提取时间对测定有明显影响,以1％Triton X-100提取18h为宜,最高比酶活达14．9U／g。在PVADH检测体系中,加入电子受体启动反应比加入酶液与底物启动反应可使酶活性分别提高60．6％和126．5％;酶液与吡咯喹啉醌（PQQ）预先保温对检测该酶活性是十分重要的,可使酶活性提高59．1％.在检测系统中加入的KCN、CaCl2和PQQ的适宜浓度分别为1．ommol／L、0．5mmol／L和2μmol／L,可使测定酶活分别提高37．1％、38．7％和214．0％．     

Preparation of polyvinyl alcohol-polyacrylamide composite polymer membrane by gamma-irradiation for entrapment of urease

Composite polymer membrane of polyvinyl alcohol (PVA) and acrylamide was prepared on cheesecloth support by gamma-irradiation induced free radical polymerization. The enzyme urease was entrapped in the membrane during polymerization and was cross-linked within the matrix using glutaraldehyde. The membranes could be reused a number of times without significant loss of urease activity.     

Phase relationships of an equivalent mixture of sulfated polyvinyl alcohol and aminoacetalyzed polyvinyl alcohol in microsalt aqueous solution

A new type of polymer salt was prepared from an equivalent mixture of partly sulfated polyvinyl alcohol (polyanion, Q^?) and partly aminoacetalyzed polyvinyl alcohol (polycation, P⁺). With respect to a three-component system composed of this polymer salt (P⁺Q^?), water, and a microsalt (K⁺A^?), phase relationships, as represented by complex coacervation, were investigated. Experimental results were discussed according to a theoretical equation for the free energy of mixing derived by taking into account the entropy and enthalpy contributions as ascribed for non-ionic polymer solution, and the electrostatic free energy expression as derived by Voorn.     

A visco-hyperelastic constitutive approach for modeling polyvinyl alcohol sponge

This study proposes the quasi-linear viscoelastic (QLV) model to characterize the time dependent mechanical behavior of poly(vinyl alcohol) (PVA) sponges. The PVA sponges have implications in many viscoelastic soft tissues, including cartilage, liver, and kidney as an implant. However, a critical barrier to the use of the PVA sponge as tissue replacement material is a lack of sufficient study on its viscoelastic mechanical properties. In this study, the nonlinear mechanical behavior of a fabricated PVA sponge is investigated experimentally and computationally using relaxation and stress failure tests as well as finite element (FE) modeling. Hyperelastic strain energy density functions, such as Yeoh and Neo-Hookean, are used to capture the mechanical behavior of PVA sponge at ramp part, and viscoelastic model is used to describe the viscose behavior at hold part. Hyperelastic material constants are obtained and their general prediction ability is verified using FE simulations of PVA tensile experiments. The results of relaxation and stress failure tests revealed that Yeoh material model can define the mechanical behavior of PVA sponge properly compared with Neo-Hookean one. FE modeling results are also affirmed the appropriateness of Yeoh model to characterize the mechanical behavior of PVA sponge. Thus, the Yeoh model can be used in future biomechanical simulations of the spongy biomaterials. These results can be utilized to understand the viscoelastic behavior of PVA sponges and has implications for tissue engineering as scaffold.     

Enzymatic methods for in situ cell entrapment and cell release

We report an enzyme-based method for the in situ entrapment of cells within a biopolymeric hydrogel matrix. Specifically, we used a calcium-independent microbial transglutaminase that is known to cross-link proteins and observed that it catalyzes the formation of gels from a pre-gel solution containing 10% gelatin and E. coli cells. Hydrogel formation occurs 2-3 h after adding transglutaminase, and no additional external intervention is required to initiate gel formation. The in situ entrapped cells grow rapidly and to high cell densities within the gelatin hydrogel. Additionally, the entrapped cells respond to isopropylthiogalactoside induction. The cross-linked gelatin network can be rapidly hydrolyzed (within 1 h) by the protease, proteinase K. Treatment of the network by this protease releases the entrapped E. coli cells. These cells appear unharmed by proteinase K; they can grow and be induced after protease treatment. The ability to in situ entrap, grow, and release cells under mild conditions provides unique opportunities for a range of applications and should be especially useful for microfluidic biosensor systems.     

Micropatterning polyvinyl alcohol as a biomimetic material through soft lithography with cell culture

Integrating inorganic materials with specific biological systems requires a complex set of properties that can mimic physiological function to create microchip devices. Several factors in the material including the spatial location of the cellular attachment heavily influence the control of this functionality and must be dictated to parallel in vivo characteristics. In this work, we demonstrate polyvinyl alcohol (PVA) for use in emulating these properties to produce a highly robust and functional biomaterial as a microchip. We show the utility of PVA for cellular patterning using soft lithography. Based on these results, PVA can be applied in a diversity of areas including tissue engineering, biomimetics, and cellular micropatterning.     

Biodegradable thermoplastic composites based on polyvinyl alcohol and algae

Algae constitute a largely available, low value material from renewable resources of marine origin to be used for the production of eco-compatible composites. Fibers of the green alga Ulva armoricana from the French coast were positively evaluated for the production of composites with a hydrophilic, eco-compatible polymer, such as poly(vinyl alcohol) (PVA) as continuous matrix by casting of aqueous suspensions and compression molding. PVA, Ulva, and starch were also successfully processed by the melt in the presence of glycerol. Positive results were obtained for film-forming properties and mechanical characteristics also with limited amounts of PVA (40%) attesting for Ulva suitability to be introduced in composites (up to 30%). Degradation in soil of Ulva and an Ulva-based composites outlined a rapid mineralization of Ulva in the selected medium (over 80% in 100 days) while the composite samples underwent a mineralization rate affected by the different component propensity to degradation.     

The effect of polyvinyl alcohol and polyvinyl pyrrolidone on diffusion artifacts in lactate dehydrogenase histochemistry

Summary The effect of polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP), alone and in combination, on diffusion artifacts in histochemical incubations has been investigated using LDH as model enzyme. By measuring the amount of formazan in the medium at the end of the incubation it has been shown that both substances, but especially PVA, are effective in limiting diffusion. The significance of this is discussed in general as well as in relation to other procedures used to reduce diffusion artifacts.The Following Abbreviations are used in the Article NAD -Nicotinamide Adenine Dinucleotide - NADH -Nicotinamide Adenine Dinucleotide, Reduced form - PVA Polyvinyl alcohol - PVP Polyvinyl pyrrolidone - PMS Phenazine methosulfate - tris tris (hydroxymethyl)-aminomethan - Nitro-BT Nitro Blue Tetrazolium - LDH Lactic dehydrogenase     

Immobilization of champagne yeasts by inclusion into cryogels of polyvinyl alcohol for preventing cell escape from carrier matrix

Wine champagnizing, a process involving the use of champagne yeasts immobilized by inclusion into cryogels of polyvinyl alcohol, has been studied. Treatment of yeast cells with the autoregulatory factor d1 was proposed as a means of preventing the cell escape from the carrier matrix. Such a treatment inhibited growth and proliferation processes in yeasts cells, without affecting the activity of fermentation; the resulting champagne had the same organoleptic and chemical characteristics as its counterparts obtained using traditional techniques.     

Some effects of polyvinyl alcohol and polyvinyl pyrrolidone on the activity of lactate dehydrogenase and its isoenzymes

Summary The activity of Sigma type II LDH (containing all LDH isoenzymes), type VII (LDH 1) and type V (LDH 5) has been evaluated in 0.1 M phosphate buffer and 0.4 M tris-HCl buffer, both at pH 7.4, before and after addition of PVP and PVA. A slight reduction of reaction velocity occurred when PVP and PVA were added, probably because of the increased viscosity of the medium. More specifically, PVA seemed to reduce the LDH 1 activity in phosphate buffer, but it seemed to counteract, together with PVP, conformational changes due to tris-HCl buffer of high molarity. None of these effects were found to influence the results of section histochemistry, but it could be shown that PVA was able to reduce urea inhibition of LDH 5 in tris-HCl buffer.The following Abbreviations are used in the Article PVA Polyvinyl alcohol - PVP Polyvinyl pyrrolidone - tris tris (hydroxymethyl)-aminomethan - LDH Lactate dehydrogenase - -NADH -Nicotinamide Adenine Dinucleotide, reduced form - PMS Phenazine methosulfate - BSA Bovine serum albumin     

Procedure for obtaining and catalytic properties of trypsin immobilized in cryogels of polyvinyl alcohol

Commercial preparations of trypsin, varying in activity, were immobilized in a cryogel of polyvinyl alcohol, activated by dialdehydes (terephthalic, succinic, or glutaric) or divinyl sulfone. All preparations of the immobilized enzyme exhibited hydrolytic activity and retained stability for 8 months. In an organic solvent environment, specimens of immobilized trypsin catalyzed the synthesis of N-carbobenzoxy-L-phenylalanyl-L-arginyl-L-leucine p-nitroanilide from N-carbobenzoxy-L-phenylalanyl-L-argininine methyl ester (or N-carbobenzoxy-L-phenylalanyl-L-arginine) and L-leucine p-nitroanilide, as well as the formation of N-carbobenzoxy-L-alanyl-L-alanyl-L-arginyl-L-phenylalanine p-nitroanilide from N-carbobenzoxy-L-alanyl-L-alanyl-L-arginine and L-phenylalanine p-nitroanilide. The presence of small amounts of water in organic solvents was prerequisite to the biocatalysts manifesting synthase activity in reactions of peptide bond formation.     

Synthesis of tolerogenic monomethoxypolyethylene glycol and polyvinyl alcohol conjugates of peptides

Recent studies from this laboratory showed that tolerogenic peptide conjugates are very effective reagents for obtaining epitope-specific immunosuppression of antibody responses to immunopathogenic sites on multideterminant complex protein antigens. This paper describes the procedure for synthesis of well-defined conjugates of peptides to monomethoxypoly-ethylene glycol (mPEG) or to polyvinyl alcohol (PVA). The first step involves succinylation of the hydroxyl groups on the polymers by reaction with succinic anhydride. The polymer is then coupled via the carboxyl of the succinyl group to the -NH₂ of the completed peptide on the synthetic resin, while maintaining intact all the side-chain protecting groups on the peptide. The mPEG or PVA-peptide conjugates are cleaved from the resin and purified by standard procedures. This method results in the preparation of conjugates in which one molecule of tolerogenic polymer is coupled to the N-terminal of an otherwise unaltered peptide molecule.     

Use of polyvinyl alcohol as a stabilizer of peroxidase-antibody conjugate for enzyme immunoassay

Summary The activity of a peroxidase-antibody conjugate was greatly stabilized while in solution through the addition of polyvinyl alcohol (PVA). This stabilizing effect was dependent upon the molecular weight and degree of hydrolysis of the PVA used. The concentrations of PVA necessary for maximum stabilization had no adverse effects upon enzyme immunoassay. Thus, the conjugate stabilized by PVA can be used without dilution for enzyme immunoassay, and without need of refrigeration during transport and storage.     

Evaluation of polyvinyl alcohol for fatty acid supplementation in adipose tissue explant culture

Cultures of adipose tissue explants are a valuable tool for studying the intracellular mechanisms involving hormones and nutrients. However, testing how fatty acids affect cells requires a carrier molecule; bovine serum albumin (BSA) has been used for this purpose. However, contaminants can alter the cellular response. Our objectives were to: 1) test BSA as a fatty acid carrier and 2) evaluate polyvinyl alcohol (PVA) as a replacement for BSA. Adipose tissue explants from nine pigs were cultured in medium 199 for 4, 12, 24, and 48 h, with the following treatments: control, PVA (100 mM PVA added) and PVA + pGH (100 mM PVA plus 0.1 mg/mL porcine growth hormone). After each culture period, explants were collected and assayed for lipogenesis. After 48 h in culture, explants were assayed for lipolysis. A preliminary study with different commercial sources and high concentrations showed that BSA affected lipogenic rates. On the other hand, there were no effects of PVA on lipid synthesis, while pGH (positive control) reduced glucose incorporation into lipids (P < 0.01) when compared to both control and PVA (P < 0.05). There was no difference between control and PVA for lipolysis rates. However, pGH increased lipolysis when compared to control (P < 0.01) and PVA (P < 0.05). We demonstrated that BSA can alter lipogenesis, which precludes its use as a carrier molecule. On the other hand, addition of PVA had no effect on lipolysis or lipogenesis. We suggest the use of PVA instead of BSA for adding bioactive fatty acids to cultures of adipose tissue.     

Stabilization of polyvinyl alcohol coating of polyester cloth for reduction of bacterial adhesion

Bacterial adhesion onto polyester cloth was greatly reduced by coating the cloth with polyvinyl alcohol (PVA). Under autoclaving conditions, the PVA coating was stable in water but not in 1% SDS. However, the treatment of the PVA-coated cloth with acidic 1% glutaraldehyde stabilized the coating to retain full resistance to bacterial adhesion even after autoclaving in 1% SDS.