Shao-Tong Jiang

Production of l -lactic acid by Rhizopus oryzae using semicontinuous fermentation in bioreactor

Semicontinuous fermentation using pellets of Rhizopus oryzae has been recognized as a promising technology for l-lactic acid production. In this work, semicontinuous fermentation of R.xa0oryzae ASxa03.819 for l-lactic acid production has been developed with high l-lactic acid yield and volumetric productivity. The effects of factors such as inoculations, CaCO3 addition time, and temperature on l-lactic acid yield and R.xa0oryzae morphology were researched in detail. The results showed that optimal fermentation conditions for the first cycle were: inoculation with 4% spore suspension, CaCO3 added to the culture medium at the beginning of culture, and culture temperature of 32–34°C. In orthogonal experiments, high l-lactic acid yield was achieved when the feeding medium was (g/l): glucose, 100; (NH4)2SO4, 2; KH2PO4, 0.1; ZnSO4·7H2O, 0.33; MgSO4·7H2O, 0.15; CaCO3, 50. Twenty cycles of semicontinuous fermentation were carried out in flask culture. l-lactic acid yield was 78.75% for the first cycle and 80–90% for the repeated cycles; the activities of lactate dehydrogenases (LDH) were 7.2–9.2xa0U/mg; fermentation was completed in 24xa0h for each repeated cycle. In a 7-l magnetically stirred fermentor, semicontinuous fermentation lasted for 25 cycles using pellets of R.xa0oryzae ASxa03.819 under the optimal conditions determined from flask cultures. The final l-lactic acid concentration (LLAC) reached 103.7xa0g/l, and the volumetric productivity was 2.16xa0g/(l·h) for the first cycle; in the following 19 repeated cycles, the final LLAC reached 81–95xa0g/l, and the volumetric productivities were 3.40–3.85xa0g/(l·h).

Effects of partial hydrolysis and subsequent cross-linking on wheat gluten physicochemical properties and structure

The rheological behavior and thermal properties of wheat gluten following partial hydrolysis using Alcalase and subsequent microbial transglutaminase (MTGase) cross-linking were investigated. The wheat gluten storage modulus (G) and thermal denaturation temperature (Tg) were significantly increased from 2.26 kPa and 54.43°C to 7.76 kPa and 57.69°C, respectively, by the combined action of partial hydrolysis (DH 0.187%) and cross-linking. The free SH content, surface hydrophobicity, and secondary structure analysis suggested that an appropriate degree of Alcalase-based hydrolysis allowed the compact wheat gluten structure to unfold, increasing the β-sheet content and surface hydrophobicity. This improved its molecular flexibility and exposed additional glutamine sites for MTGase cross-linking. SEM images showed that a compact 3D network formed, while SDS-PAGE profiles revealed that excessive hydrolysis resulted in high-molecular-weight subunits degrading to smaller peptides, unsuitable for cross-linking. It was also demonstrated that the combination of Alcalase-based partial hydrolysis with MTGase cross-linking might be an effective method for modifying wheat gluten rheological behavior and thermal properties.

Transglutaminase-induced gelation properties of soy protein isolate and wheat gluten mixtures with high intensity ultrasonic pretreatment.

Soy protein isolate (SPI) and wheat gluten (WG) are widely used in commercial food applications in Asia for their nutritional value and functional properties. However, individually each exhibits poor gelation. In this study, we examined the microbial transglutaminase (MTGase)-induced gelation properties of SPI and WG mixtures with high intensity ultrasonic pretreatment. Ultrasonic treatment reduced the particle size of SPI/WG molecules, which led to improvements in surface hydrophobicity (Ho) and free sulfhydryl (SH) group content. However, MTGase crosslinking facilitated the formation of disulfide bonds, markedly decreasing the content of free SH groups. Ultrasonic treatment improved the gel strength, water holding capacity, and storage modulus and resulted in denser and more homogeneous networks of MTGase-induced SPI/WG gels. In addition, ultrasonic treatment changed the secondary structure of the gel samples as determined by Fourier transform infrared spectroscopic analysis, with a reduction in α-helices and β-turns and an increase in β-sheets and random coils. Thus, ultrasound is useful in facilitating the gelation properties of MTGase-induced SPI/WG gels and might expand their utilization in the food protein gelation industry.

Effects of microwave pretreatment and transglutaminase crosslinking on the gelation properties of soybean protein isolate and wheat gluten mixtures

BACKGROUNDnThe integration of soybean protein isolate (SPI) with wheat gluten (WG) crosslinked via microbial transglutaminase (MTGase) may enhance the formation of ϵ-(γ-glutamyl)lysine covalent bonds, because SPI is rich in lysine and WG contains more glutamine. Microwave pretreatment may accelerate enzymatic reactions. In this study, we aimed to elucidate the effects of microwave pretreatment on the gelation properties of SPI and WG crosslinked with MTGase.nnnRESULTSnInterestingly, the gel strength, water-holding capacity (WHC) and storage modulus (G) values of MTGase-induced SPI/WG gels were dramatically improved with increasing microwave power. Moreover, the MTGase crosslinking reaction promoted the formation of disulfide bonds, markedly reducing the free SH group and soluble protein content of gels. Fourier transform infrared spectroscopic analysis of SPI/WG gels showed that microwave pretreatment increased the proportion of α-helices and β-turns and decreased the proportion of β-sheets. Results from scanning electron microscopy indicated that the MTGase-induced SPI/WG gels had denser and more homogeneous microstructures after microwave pretreatment.nnnCONCLUSIONnThe effect of microwave pretreatment is useful in advancing gelation characters of MTGase-induced SPI/WG gels and provides the possibility for expanding the application of food protein.

Characterization of Wheat Germ Oil in Terms of Volatile Compounds, Lipid Composition, Thermal Behavior, and Structure

In this study, the volatile compounds, lipid composition, thermal behavior, and structure of wheat germ oil were analyzed. Forty-four volatile compounds, mainly alcohols, esters, alkenes, and aldehydes, were isolated by headspace solid phase micro extraction. The major constituents were found to be hexanal (15.97%), 2-methyl-2-butene (10.43%), 2,4-heptadienal (8.53%), and limonene (6.83%). The oil was rich in unsaturated fatty acid (83.45%), especially in linoleic acid (64.82%), and there were 75.49% unsaturated fatty acid distributed in Sn-2 position. The main content of unsaponifiable matters was sterol, especially β-sitosterol (64.64%). The thermogravimetric analysis results showed one major regime of weight loss over a temperature range of 280–500°C and an exothermic behavior was observed by the differential scanning calorimetry data.

Significant damage-rescuing effects of wood vinegar extract in living Caenorhabditis elegans under oxidative stress

BACKGROUNDnWood vinegar (WV), a byproduct from the charcoal production process, has been reported to have excellent antioxidant capability by chemical examination. However, the biological effect of WV in living animals is still unknown. In this study, a simple model organism, the nematode Caenorhabditis elegans, was used as an in vivo system to assess the biological effects of wood vinegar through the development, lifespan, brood size, germline cell apoptosis and superoxide dismutase (SOD) level.nnnRESULTSnWood vinegar extract (WVE) promoted the development, prolonged the lifespan and increased the brood size in reactive oxidative species (ROS)-sensitive mutant worms. WVE treatment rescued the effects of damage in germline cell apoptosis and SOD upregulation induced by paraquat, an ROS generator, to the control level. Additionally, WVE showed comparative ability in rescuing damage as compared with L-ascorbic acid and α-tocopherol.nnnCONCLUSIONnWVE treatment exhibits a remedial/beneficial effect on ROS-sensitive mutant under normal cultural conditions and on wild-type worms under oxidative stress. ROS scavenging is involved in the damage-rescuing mechanism. This study will provide a basal biological and nutritional exploration for the use of WV as a functional food, and for the substitution of chemical antioxidants with side effects in food.

Effect of Modified Wheat Gluten on Boiling Resistance Capacity of Pork Meatballs.

The effect of the modified wheat gluten (MWG) extender, prepared by alcalase-based hydrolysis and transglutaminase cross-linking, on meatballs was analyzed in this study. Here, we studied the effect of MWG addition on the boiling resistance capacity of pork meatballs (MB-MWG) at high temperature (100 °C) and increasing cooking time; meatballs with added soy protein isolates (MB-SPI) and raw wheat gluten (MB-WG) were used as references. The cooking loss, water-holding capacity (WHC), and textural properties of meatballs were investigated. The results revealed that MB-MWG showed lower cooking loss, which decreased by 49.16% compared to meatballs without added extenders when treated for 30 min. The WHC of MB-MWG significantly increased from 80.68% to 95.42%. The hardness, springiness, and chewiness (textural properties) of MB-MWG were also significantly increased by 97.05%, 6.68%, and 121.96%, respectively. The addition of MWG increased the cross-linking in meatballs during the cooking process, as indicated by the higher G. SDS-PAGE indicated an obvious decrease in myosin heavy chain in MB-MWG cooked for 30 min at 100 °C, which was attributed to the interaction of myofibrillar proteins in pork meat with MWG. The nuclear magnetic resonance T2 relaxation time patterns indicated that MWG addition caused an increase in the bound water content, and decrease in the free water content, of meatballs. An analysis of the microstructures revealed that the MB-MWG formed the most regular and compact network. Therefore, MWG could be used as an ingredient to facilitate the processing of meat products.

Species diversity of Lachnum (Helotiales, Hyaloscyphaceae) from temperate China

Twenty-three temperate China species of Lachnum, Lachnum abnorme, L. angustum, L. brevipilosum, L. calosporum, L. calyculiforme, L. carneolum, L. ciliare, L. controversum, L. flavidulum, L. cf. fushanese, L. indicum, L. kumaonicum, L. lushanese, L. minutum, L. montanum, L. cf. pteridophyllum, L. pygmaeum, L. sclerotii var. sclerotii, L. sclerotii var. sichuanense, L. subpygmeaum, L. tenuissimum, L. virgineum and L. willisii are reported, whose main characteristics are given in a formula of the described species, some of which are discussed below.

Transglutaminase-set colloidal properties of wheat gluten with ultrasound pretreatments

The low solubility of wheat gluten limits its accessibility. This work aimed to study the impact of ultrasonic pretreatments on the gelation of wheat gluten. The pretreatments included ultrasound combined with alkali, urea, Na2SO3, with or without the addition of transglutaminase (TGase). The gel strength of wheat gluten was 287g/cm2 after treatment with Na2SO3/ultrasound/TGase. The free sulfhydryl and disulfide bond content was significantly affected by ultrasound treatment. After treatments including TGase crosslinking, the molecular weight of wheat gluten complexes became larger. The network formed by the wheat gluten was transformed into a dense and homogenous structure after the pretreatment with Na2SO3/ultrasound/TGase. The content of random coil of wheat gluten increased. The gelation of wheat gluten could also be significantly enhanced by Na2SO3/ultrasound treatment followed by TGase treatment. Using physical and chemical pretreatments to allow TGase to enhance the gelation of wheat gluten may increase its uses as a food additive.

Heterologous signal peptides-directing secretion of Streptomyces mobaraensis transglutaminase by Bacillus subtilis

Microbial transglutaminase (MTG) from Streptomyces mobaraensis has been widely used for crosslinking proteins in order to acquire products with improved properties. To improve the yield and enable a facile and efficient purification process, recombinant vectors, harboring various heterologous signal peptide-encoding fragments fused to the mtg gene, were constructed in Escherichia coli and then expressed in Bacillus subtilis. Signal peptides of both WapA and AmyQ (SPwapA and SPamyQ) were able to direct the secretion of pre-pro-MTG into the medium. A constitutive promoter (PhpaII) was used for the expression of SPwapA-mtg, while an inducible promoter (Plac) was used for SPamyQ-mtg. After purification from the supernatant of the culture by immobilized metal affinity chromatography and proteolysis by trypsin, 63.0u2009±u20090.6xa0mg/L mature MTG was released, demonstrated to have 29.6u2009±u20090.9xa0U/mg enzymatic activity and shown to crosslink soy protein properly. This is the first report on secretion of S. mobaraensis MTG from B. subtilis, with similar enzymatic activities and yields to that produced from Escherichia coli, but enabling a much easier purification process.