A Simple Technique for Direct Immobilization of Target Enzymes from Cell Lysates Based on the SpyTag/SpyCatcher Spontaneous Reaction.

Many of the current methods for enzyme purification and immobilization suffer from several drawbacks, such as requiring tedious multistep procedures or long preparation, and being environmentally unfriendly, due to the chemicals and conditions involved. Thus, a simple technique for direct purification and immobilization of target enzymes from cell lysates was proposed. The elastin-like polypeptides (ELPs)-SpyCatcher chimera could mediate the formation of silica carriers within seconds and the target enzymes were then covalently immobilized on silica carriers via SpyCatcher/SpyTag spontaneous reaction. These tailor-made carriers were easily prepared, with precisely controlled morphology and size, as well as none-consuming surface modification needed, which could specifically immobilize the SpyTag-fused target enzymes from the cell lysate without pre-purification.

[Analysis of differential protein expression of REMI mutant of Trichoderma koningii].

On purpose of expounding the phenotype change of Trichoderma mutant which produces cellulase, protein profiles of T. koningii CICC and its mutant TK-2R-1 were detected by 2-DE. Eight unique spots were found and four of them were identified and their functions by MS-TOF-TOF were connected with growth and metabolization of Trichoderma spp. The research provides a new theoretical basis and methods for the construction of Trichoderma mutant which is used to produce efficient cellulase.

[Effect and mechanism on dopamine contents of striatum in rat model of Parkinson's disease ginsenoside Rg1].

OBJECTIVE: To investigate effect and mechanisms on dopamine contents of striatum (Str) in the 6-OHDA induced rat model of Parkinson's disease (PD) by ginsenoside Rg1. METHOD: Ovariectomized PD rats were treated with vehicle, ginsenoside Rg1, (10 mg x kg(-1)) or estrogen receptor (ER) antagonist ICI 182,780 for 14 d. The change of apomorphine-linduced rotational behavior in PD rats were observed. The high performance lipid chromotophotography (HPLC) was used to determine the contents of DA, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA)in striatum. RESULT: Rg1 treatment could ameliorate the PD rat's rotational behavior induced by apomorphine (P < 0.01). This effect could be blocked by ER antagonist ICI 182,780. The DA, DOPAC and HVA levels in the injured side of Str for PD rats were significantly decreased compared with the intact side (P < 0.01). Rg1, treatment could increase DA contents in the injured side of Str (P < 0.01). ICI 182,780 could completely block the neuroprotective effects of Rg1. The DA contents and its metabolites in the injured side of the ICI treatment group were significantly decreased compared with the Rg1 group (P < 0.01). CONCLUSION: Ginsenoside Rg1 may have protective effects on the dopaminergic neurons for the 6-OHDA induced OVX rat model of PD, ER. May be involved in the protection action.

[The protective effect of ginsenoside Rg1 on dopaminergic neurons of substantia in the ovariectomized rat model of Parkinson's disease].

AIM: To investigate the neuroprotective effect of ginsenoside Rg1 on dopaminergic neurons of substantia nigra in ovariectomized rat model of Parkinson's disease and the possible mechanisms. METHODS: Wistar female rats were ovariectomized and treated with vehicle, ginsenoside Rg1 or 17-beta estradiol intracerebroventricularly in the 6-OHDA induced rat model of Parkinson's disease. Immunohistochemistry was used to detect the tyrosine hydroxylase (TH) immunoreactive neurons and the protein expression of Bcl-2. Perls' iron staining was used to determine the changes of iron in substantia nigra (SN). RESULTS: 910 Rg1 or 17-beta estradiol treatment could ameliorate the rat's rotational behavior induced by apomorphine. 92) Rg1 or 17-beta estradiol treatment could increase TH immunoreactive neurons in the injured side of SN compared to the 6-OHDA group. (3) Iron staining in the injured side of SN was significantly increased comparing with the contralateral side in the 6-OHDA group. Rg1 or 17-beta estradiol treatment could reverse the increase of iron staining. (4) Both Rg1 and 17-beta estradiol treatment could increase Bcl-2 protein expression in the injured side of SN compared to the 6 OHDA group. CONCLUSION: Ginsenoside Rg1 has estrogen-like activities and has neuroprotective effects on the dopaminergic neurons in the 6-OHDA induced ovariectomyzed(OVX) rat model of Parkinson's disease (PD). This effect may be attributed to attenuating iron overload and anti-apoptosis.

Neuroprotective effects of genistein on dopaminergic neurons in the mice model of Parkinson's disease.

Emerging evidence suggests beneficial effects of estrogen and estrogen-like chemicals on neurodegenerative diseases, especially Parkinson's disease (PD). Genistein, an isoflavone naturally found in soy products, displays estrogenic properties. The present study aims to investigate the neuroprotective effects of genistein on dopaminergic neurons in ovariectomized (OVX), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD model mice. MPTP significantly decreased the levels of dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striatum, which could be restored by genistein or estrogen pretreatment. MPTP-challenge with genistein or estrogen pretreatment demonstrated reduced neurotoxicity, with tyrosine hydroxylase-immunoreactive (TH-IR) neurons in the substantia nigra pars compacta (SNpc) affected to a significantly lesser extent as compared to the MPTP treated control. The reverse transcription-PCR results also confirmed that the MPTP-induced downregulation of TH, dopamine transporter (DAT) and Bcl-2 mRNA expression in the midbrain could be restored by genistein or estrogen pretreatment. These findings provide the first evidence that genistein has neuroprotective effects on dopaminergic neurons in the MPTP-induced PD mice and this effect may be attributed to enhancing Bcl-2 gene expression.