Optimization of liquid fermentation conditions for biotransformation zein by Cordyceps militaris 202 and characterization of physicochemical and functional properties of fermentative hydrolysates

Abstract Cordyceps militaris 202 is a potential fungus for biotransformation zein, due to its various proteases, high tolerance and viability in nature. In this article, single factor experiment and response surface methodology were applied to optimize the liquid fermentation conditions and improve the ability of biotransformation zein. The optimized fermentation conditions were as follows: inoculum concentration of 19%, volume of liquor of 130 mL/500 mL and pH of 4.7. Under this condition, the degree of hydrolysis (DH) was 27.31%. The zein hydrolysates from fungi fermentation maintained a high thermal stability. Compared to the original zein, the zein hydrolysates were found to have high solubility, which most likely results in improved foaming and emulsifying properties. Overall, this research demonstrates that hydrolysis of zein by C. militaris 202 is a potential method for improving the functional properties of zein, and the zein hydrolysates can be used as functional ingredients with an increased antioxidant effect in both food and non-food applications.

In vitro and in vivo evaluation of the developed PLGA/HAp/Zein scaffolds for bone-cartilage interface regeneration / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the effect of electronspun PLGA/HAp/Zein scaffolds on the repair of cartilage defects.</p><p><b>METHODS</b>The PLGA/HAp/Zein composite scaffolds were fabricated by electrospinning method. The physiochemical properties and biocompatibility of the scaffolds were separately characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), and fourier transform infrared spectroscopy (FTIR), human umbilical cord mesenchymal stem cells (hUC-MSCs) culture and animal experiments.</p><p><b>RESULTS</b>The prepared PLGA/HAp/Zein scaffolds showed fibrous structure with homogenous distribution. hUC-MSCs could attach to and grow well on PLGA/HAp/Zein scaffolds, and there was no significant difference between cell proliferation on scaffolds and that without scaffolds (P>0.05). The PLGA/HAp/Zein scaffolds possessed excellent ability to promote in vivo cartilage formation. Moreover, there was a large amount of immature chondrocytes and matrix with cartilage lacuna on PLGA/HAp/Zein scaffolds.</p><p><b>CONCLUSION</b>The data suggest that the PLGA/HAp/Zein scaffolds possess good biocompatibility, which are anticipated to be potentially applied in cartilage tissue engineering and reconstruction.</p>

Effect of shuanghuangbu on the adhesion growth and proliferation of human periodontal ligament cells on the zein stent / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Shuanghuangbu on the adhesion growth and proliferation of human periodontal ligament cells (HPDLCs) on the zein scaffold, and to study the feasibility of Shuanghuangbu zein used in the periodontal tissue engineering as a compound scaffold material.</p><p><b>METHODS</b>HPDLCs were cultured in vitro using the explants adherent method. Shuanghuangbu Decoction was prepared using water extraction and alcohol precipitation method. Zein scaffold was prepared using solvent casting/corpuscle leaching method, which was dispensed into Shuanghuangbu culture fluid at the concentration of 50, 100, 150, 200, 500, 1000 microg/mL, respectively. Shuanghuangbu of different concentrations groups, the zein scaffold leaching liquid group, and the cell control group were also set up. The effect of each group on the proliferation of HPDLCs was detected by MTT. According to results of MTT, the best concentration of Shuanghuangbu (100 microg/mL) was selected as the experiment group. Ten percent FBS of DMEM culture fluid was taken as the control group. The effect of Shuanghuangbu on the morphology of HPDLCs on the zein scaffold was observed using inverted phase contrast microscope and scanning electron microscope.</p><p><b>RESULTS</b>Compared with the zein scaffold leaching liquid group and the cell control group, the proliferation of HPDLCs could be promoted by different concentrations of Shuanghuangbu culture fluid, with the most obvious effect shown by Shuanghuangbu at 100 microg/mL (P<0.01). There was no obvious difference between the zein scaffold leaching liquid group and the cell control group (P>0.05), but their cell proliferation rates were positive values. The morphological results demonstrated that HPDLCs successfully grew on the zein scaffold. The number of HPDLCs under the action of Shuanghuangbu was more and the morphous was better.</p><p><b>CONCLUSIONS</b>Shuanghuangbu could promote the adhesion growth and proliferation of HPDLCs on the zein scaffold. Shuang- huangbu zein was feasible to be used in the periodontal tissue engineering as a compound scaffold materia.</p>

The use of zein and Shuanghuangbu for periodontal tissue engineering / 国际口腔科学杂志·英文版

<p><b>AIM</b>Tissue engineering is a promising area with a broad range of applications in the fields of regenerative medicine and human health. The emergence of periodontal tissue engineering for clinical treatment of periodontal disease has opened a new therapeutic avenue. The choice of scaffold is crucial. This study was conducted to prepare zein scaffold and explore the suitability of zein and Shuanghuangbu for periodontal tissue engineering.</p><p><b>METHODOLOGY</b>A zein scaffold was made using the solvent casting/particulate leaching method with sodium chloride (NaCl) particles as the porogen. The physical properties of the zein scaffold were evaluated by observing its shape and determining its pore structure and porosity. Cytotoxicity testing of the scaffold was carried out via in vitro cell culture experiments, including a liquid extraction experiment and the direct contact assay. Also, the Chinese medicine Shuanghuangbu, as a growth factor, was diluted by scaffold extract into different concentrations. This Shuanghuangbu-scaffold extract was then added to periodontal ligament cells (PDLCs) in order to determine its effect on cell proliferation.</p><p><b>RESULTS</b>The zein scaffold displayed a sponge-like structure with a high porosity and sufficient thickness. The porosity and pore size of the zein scaffold can be controlled by changing the porogen particles dosage and size. The porosity was up to 64.1%-78.0%. The pores were well-distributed, interconnected, and porous. The toxicity of the zein scaffold was graded as 0-1. Furthermore, PDLCs displayed full stretching and vigorous growth under scanning electronic microscope (SEM). Shuanghuangbu-scaffold extract could reinforce proliferation activity of PDLCs compared to the control group, especially at 100 microg x mL(-1) (P < 0.01).</p><p><b>CONCLUSION</b>A zein scaffold with high porosity, open pore wall structure, and good biocompatibility is conducive to the growth of PDLCs. Zein could be used as scaffold to repair periodontal tissue defects. Also, Shuanghuangbu-scaffold extract can enhance the proliferation activity of PDLCs. Altogether, these findings provide the basis for in vivo testing on animals.</p>

Relationship between structural and biochemical characteristics and texture of corn grains

The texture of corn grains is a fundamental characteristic for the industry as well as for grain producers and processors. To further understand the mechanisms involved in grain hardness, contrasting corn cultivars for grain hardness and protein quality were evaluated. The cultivars were Cateto L237/67 (hard endosperm and low protein value), QPM BR 451 (semi-hard endosperm and high protein value); Bolivia-2 (floury endosperm and low protein value), and Opaque-2 (floury endosperm and high protein value). Evaluations were carried out at 10, 20, 30, 40, 50, and 60 days after pollination in developing corn grains and in the mature grain. In developing grains, evaluation consisted in the determination of the area, percentage of starch granules, distribution of starch granules, and protein bodies in the endosperm. In mature corn grains, the parameters evaluated were: density, percentage of total proteins, levels of lysine and tryptophan, and banding pattern of zeins. The results indicate that the higher physical resistance of corn grains from the cultivars analyzed is influenced by the high percentage of total proteins, high synthesis of 27-kDa zeins, presence of protein bodies, and perfect organization of starch granules in the endosperm, independent of their sizes.

Extracción y caracterización de las zeínas del grano de diez cultivares de maíz / Extraction and characterization of zeins from kernels of 10 maize cultivars

The study consisted in the extraction and characterization of zeins of Venezuelan maizes, according to their solubility and molecular properties. The cultivars analyzed were the hybrids Ceniap PB8, Tocoron 127, Arichuna, Obregón, Ceniap 3, Corocito 101 and the varieties Máquina del Ceniap, Venezuela-1 and Venezuela-1 Opaco-2. Zeins were extracted with 70 ethanol and fractionated by two methods: 95 ethanol and column filtration with Sephadex G-200. The molecular weight was determined by electrophoresis in a discontinuous polyacrilamide gel with sodium dodecyl sulfate (PAGE-SDS). The results demonstrated that zeins account for 36.57 of the total protein present in normal corn grain and 9.38 in Opaque-2 corn. Prolamines presented a soluble fraction in 95 ethanol (alpha zeins) which represented 33,12 of zeins and another insoluble (beta zeins) the 66.88. In column fractionation, three fractions were obtained, two major A and C and another B in minor quantity, which varied in proportion between cultivars. Zeins which were reduced with 2-mercaptoethanol separated into two subunits with molecular weights of 21,300 and 24,300 daltons, whereas unreduced zeins presented large sized aggregates which remained at the origin and large numbers of bands (nine) whose molecular weights oscillated between 24,300 and 87,000 daltons. The electrophoretic patterns of normal maizes were similar, but differ from the pattern of the Opaque-2. In this maize, zeins reduced presented only the band with molecular weight 24,300 daltons and the unreduced zeins showed those with molecular weights of 87,000; 78,500; 48,500 and 24,300 daltons. Reduced alpha and beta are made up of the same basic components of zeins. Likewise unreduced alpha zeins are made up of the same polypeptides as unreduced zeins. The difference between Opaque-2 zeins and normal ones in that they contain more alpha zeins and less polypeptides of high molecular weight.