Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Effect of Different Fertilization Methods on Soil Biological Indexes

Fertilization plays an important role in crop growth and soil improvement. This study was conducted to determine the best fertilization system for wheat production. Experiments were arranged in a complete block design with three replications in two years. Main plots consisted of six methods of fertilization including (N1): farmyard manure; (N2): compost; (N3): chemical fertilizers; (N4): farmyard manure + compost; (N5): farmyard manure + compost + chemical fertilizers and (N6): control were arranged in sub plots. The addition of compost or farm yard manure significantly increased the soil microbial biomass carbon in comparison to the chemical fertilizer. The dehydrogenase, phosphatase and urease activities in the N3 treatment were significantly lower than in the farm yard manure and compost treatments.

Effect of Ginger and L-Carnitine on the Reproductive Performance of Male Rats

In this study, we investigated the effects of ginger and L-carnitine on the reproductive performance of male rats with respect to semen parameters, male sex hormones and the testicular antioxidant system. A total of sixty mature male albino rats were divided into four groups of fifteen rats. The control group received saline, whereas the other three groups received ginger (100 mg kg-1 d- 1.), L-carnitine (150 mg kg-1 d-1.) or a combination of both ginger (100 mg kg-1 d-1.) and L-carnitine (150 mg kg-1 d-1.) via a stomach tube daily for one month. At the end of the treatment period, the rats were sacrificed, and their sperm characteristics (count, motility and viability), antioxidant enzyme factors levels (reduced glutathione, catalase, superoxide dismutase and total antioxidant capacity) and sex hormone levels (testosterone, Follicle stimulating hormone(FSH) and luteinizing hormone (LH) were analysed. Our results showed that the three experimental treatments improved sperm parameters, antioxidant enzyme activity and testosterone hormone levels; the most pronounced positive effects were observed in the group that received a combination of both ginger and L-carnitine. Therefore, the administration of a combination of ginger and L-carnitine may be beneficial for improving male sexual performance.

The Effectiveness of Ultrasound Treatment on the Germination Stimulation of Barley Seed and its Alpha-Amylase Activity

In the present study, the effects of ultrasound as emerging technology were investigated on germination stimulation, amount of alpha-amylase activity on dry barley seeds before steeping stage of malting process. All experiments were carried out at 20 KHz on the ultrasonic generator in 3 different ultrasonic intensities (20, 60 and 100% setting from total power of device) and time (5, 10 and 15 min) at constant temperature (30C). For determining the effects of these parameters on enzyme the Fuwa method assay based on the decreased staining value of blue starch–iodine complexes employed for measurement an activity. The results of these assays were analyzed by Qualitek4 software using the Taguchi statistical method to evaluate the factor-s effects on enzyme activity. It has been found that when malting barley is irradiated with an ultrasonic power, a stimulating effect occurs as to the enzyme activity.

Improvement of Lipase Catalytic Properties by Immobilization in Hybrid Matrices

Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.