Abstract:As thermostable enzymes and organisms are much more needed, researches on heat shock proteins(HSPs) of hyperthermophilic archaea have drawn more concerns. HSPs from hyperthermophilic archaea are concise only with HSP60, sHSP, prefoldin and AAA+proteins, but without HSP100s, HSP90s, HSP70 (DnaK), HSP40 (DnaJ) and GrpE which are common in mesophilic or thermophilic archaea. Accordingly, studies on the structure, function and operation mechanism of these four groups are much more important and meaningful. This review focuses on the recent progress in the researchs on the structure, function, operation mechanism and cooperation of the HSPs from hyperthermophilic archaea. The problems and obfuscations in these HSPs are analyzed, and farther research direction and key points are put out.

Abstract:Practically all organic chemicals and plastics are nowadays produced from crude oil and natural gas. However, it is possible to produce a wide range of bulk chemicals from renewable resources by application of biotechnology. This paper focuses on White Biotechnology, which makes use of bacteria (or yeasts) or enzymes for the conversion of the fermentable sugar to the target product. It is shown that White Biotechnology offers substantial savings of non-renewable energy use and greenhouse gas emissions for nearly all of the products studied. Under favorable boundary conditions up to two thirds (67%) of the current non-renewable energy use for the production of the selected chemicals can be saved by 2050 if substantial technological progress is made and if the use of lignocellulosic feedstocks is successfully developed. The analysis for Europe (EU 25 countries) shows that land requirements related to White Biotechnology chemicals are not likely to become a critical issue in the next few decades, especially considering the large unused and underutilized resources in Eastern Europe. Substantial macroeconomic savings can be achieved under favourable boundary conditions. In principle, natural bacteria and enzymes can be used for White Biotechnology but, according to many experts in the fields, Genetically Modified Organisms (GMO) will be necessary in order to achieve the high yields, concentrations and productivities that are required to reach economic viability. Safe containment and inactivation of GMOs after release is very important because not all possible implications caused by the interaction of recombinant genes with other populations can be foreseen. If adequate precautionary measures are taken, the risks related to the use of genetically modified organisms in White Biotechnology are manageable. We conclude that the core requirements to be fulfilled in order to make clear steps towards a bio-based chemical industry are substantial technological progress in the bioprocess step and in downstream processing, high prices for fossil fuels and low prices for fermentable sugar. We strongly recommend to develop an integrated White Biotechnology strategy taking into account these four core requirements and other important accompanying activities.

Abstract:Gene expression in rice roots under nutritional stress was studied using micro array techniques. The results showed that when re-supplied with sufficient amounts of nutrition after nutrition stress, the expression of OsPra2 (a small G protein which is homologous with Pea Pra2 protein) decreased in the plants root tissue. The cDNA sequence of the OsPra2 gene and its promoter, which is about 1 kb upstream of the translation origin point, was obtained using RT-PCR and PCR approaches. The OsPra2 protein contains four conserved GTP/GDP binding domains and specific domain of Rab small G protein family. The expression of OsPra2 and GST fusion protein in onion epidermal cells showed that OsPra2 protein was localized in the membrane and nucleus of the cell. The fusion expression of OsPra2 promoter and GUS reporter gene in transgenic rice suggested that the OsPra2 promoter allowed GUS expression in coleoptiles and roots. Compared with wild type rice, OsPra2 over expressed transgenic rice showed an obvious dwarf phenotype which resembles the BR deficient rice.

Abstract:We first cloned the dsz operon of Pseudomonas delafieldii R-8 into the expressing plasmid (pPR9TT) to construct the recombinant plasmid pPR-dsz, and then reintroduced it into strain R-8 to obtain a muti-copy dsz operon engineering strain R-8-1. Compared with the wild-type, strain R-8-1 showed a higher desulfurization activity for dibenzothiophene (DBT). Initial rates of DBT removal by strain R-8-1 were 6.25 mmol/g dry cell/h, about 2-fold higher than that for wild-type strain. The recombinant cells were also applied in the desulfurization of diesel. It resulted in a 68% reduction of total sulfur from 310.8 mg/L to 100.1 mg/L, whereas only 53% of sulfur was removed by strain R-8. The stability of pPR-dsz in strain R-8-1 was studied. The results revealed the first obtain a muti-copy dsz operon engineering strain are helpful for further development in biodesulfurization.

Abstract:The full length cDNA of silkworm hibadh gene was cloned by RT-PCR and RACE (Rapid amplification of cDNA ends) technique. The hibadh gene and its deduced amino acid sequences were analyzed. The tissue distribution of hibadh gene in 5th instar silkworm larvae was tested by RT-PCR. The expression patterns of hibadh gene in simulated weightless environment were analyzed by real time RT-PCR. The results showed that the full length hibadh cDNA sequence was 1074 bp in lenth, including an open read frame of 969 bp encoding the entire coding region of Hibadh (GenBank accession No. EU719652). The deduced amino acid sequence similarities of hibadh between silkworm and Burkholderia ambifaria, Drosophila melanogaster, Apis mellifera, Xenopus tropicalis, Mus musculus, Homo sapiens were 46%, 43%, 48%, 44%, 45%, 45%, respectively. Signal peptide analysis showed that Hibadh was a secretory protein. There wasn’t glycosyl-phosphatidyl inositol anchor site in Hibadh amino acid sequence. Molecular weight and isoelectric point of Hibadh were 34.1 kD and 9.14 respectively. The RT-PCR tests indicated that the hibadh gene expressed in head, silk gland, midgut, cuticle, blood, fat body, tuba malpighii of the 5th instar silkworm larvae. There were different expression patterns of hibadh gene during different silkworm embryo period in simulated weightless environment. Simulated weightlessness resulted in the expression of silkworm hibadh gene up regulated 2.3-fold (P<0.05), up regulated 4.6-fold (P<0.01), down regulated 7.6-fold (P<0.01), down regulated 2.6-fold (P<0.05) during apophysis formation period, inverse period, trachea formation period, and whole embryo period, respectively. There was no significant change of hibadh gene expression during other period of silkworm embryo between simulated weightless and control groups. There were different response patterns to simulated weightless environment between hibadh gene and whole body of silkworm. Gene showed much higher sensitivity compared to whole body in response to environment. This study is useful for the further research on the gravity biological mechanism of hibadh gene.

Abstract:To produce recombinant Maxadilan using gene engineering technology, the gene of recombinant Maxadilan which expressed in protocaryon were designed and synthesized according to the amino acid sequences of Maxadilan. The recombinant plasmid pKYB-MAX was constructed and transformed into host bacteria Escherichia coli strain ER2566. After the MAX-intein-CBD fusion protein was purified by chintin-affinity chromatography, the self-cleavage activity of the intein was induced by β-mercaptoethanol and the recombinant Maxadilan was released from the chitin-bound intein tag. The molecular weight of peptides was determined by the laser flight mass spectrometry and the results was conformity with the theoretical value. The biological activity analysis showed that recombinant Maxadilan significantly enhanced the concentrat ion of serum glucose.

Abstract:Taking Matou goat ear margin as the study material, we succeeded in established a fibroblast cell line by the method of explant culture directly. Observations on morphology, dynamic growth, determination of viability, analysis of karyotype, test of microorganism and other characteristics were detected. Results showed: Population Doubling Time (PDT) of cells was approximately 36 h; Cell viability was 96.7% after thawing; The status of cell After passage was constant; Analysis of chromosomal karyotyps indicated that diploid (2n=60) account for 98% in the cell line. Every index in the cell line met all the standard quality controls of ATCC in USA. The established of Matou goat ear fibroblast cell line has not only important genetic resources preserved at the cell level, but also valuable material for genome, postgenome and somatic cell nuclear transfer research.

Abstract:We transfect human neural stem cells using lentiviral vectors encoding glial cell line derived neurotrophic factor (GDNF) to study its expression level in vitro and to get a stable cell line expressing GDNF. First, GDNF gene was sub-cloned into the lentiviral transfer vectors. Then, the recombinant lentiviral supernatants were packaged by 293T cells through three plasmids transient co-transfection method using standard lipofectamine reagent. The viral titers were tested by the transfection efficiency of 293T cells. At the same time, hNSChuman neural stem cells were transfected under different multiplicity of infection. GDNF gene expression level and its protein secretion level of hNSC were tested by real-time PCR and ELISA methods after transfection. Lentiviral vectors encoding GDNF were constructed. Using lentiviral vectors encoding GDNF we successfully transfected human neural stem cells, and got a stable neural stem cell lines over-expressing GDNF. Furthermore, the results indicated that GDNF expression was influenced by the multiplicity of infection. Human neural stem cells could over-express GDNF through lentivial vectors tranfection. Its gene expression level and protein expression level correlate with the multiplicity of infection.

Abstract:The effect of lignin containing natural substrates corn-cob, coir-dust, saw-dust, wheat straw and bagasse particles on the extracellular secretion of laccase in the liquid culture growth medium of Pleurotus sajor-caju MTCC 141 has been studied. The culture conditions for maximum secretion of laccase by Pleurotus sajor-caju MTCC 141 have been optimized. Homogeneous preparation of laccase from the culture filtrate of the fungus has been achieved using ammonium sulphate precipitation, anion exchange chromatography on DEAE and gel filtration chromatography on Sephadex G-100. The purified enzyme preparation gave a single protein band in SDS-PAGE analysis indicating a molecular weight of 90 kD. The enzymatic characteristics Km, kcat, pH and temperature optima of the purified laccase have been determined using 2, 6-dimethoxyphenol as the substrate and have been found to be 35 mmol/L, 0.30 min-1, 4.5oC and 37oC respectively. The Km values for the other substrate like catechol, m-cresol, pyrogallol and syringaldazine have also been determined which were found to be 216 mmol/L, 380 mmol/L, 370 mmol/L and 260 mmol/L respectively.

Abstract:Curtobacterium luteum, a gram-positive psychrotrophic bacterium, secreting an extracellular protease was isolated from the soil of Gangotri glacier, Western Himalaya. The maximum enzyme production was achieved when isolate was grown in a pH-neutral medium containing skim milk at 15oC over 120 hour. The metal ions such as Zn2+ and Cr2+ enhanced enzyme production. The specific activity of purified enzyme was 8090 u/mg after 34.1 fold purification. The 115 kD enzyme was a metalloprotease (activity inhibited by EDTA and EGTA) and showed maximum activity at 20oC and pH 7. The enzyme was active over a broad pH range and retained 84% of its original activity between pH 6–8. There was no loss in enzyme activity when exposed for 3 hours at 4oC-20oC. However, lost 65% of activity at 30oC, and was almost inactivated at 50oC, but was resistant to repeated freezing and thawing. The enzyme activity was stimulated by manganese ions; however, it was inactivated by copper ions.

Abstract:Based on carbon metabolic pathway analysis of Escherichia coli MG1655, an aerobic succinate fermentation platform was constructed by knocking out five genes (ptsG, poxB, pta, iclR and sdhA), which was named E. coli QZ1111. Flask cultivation results showed that E. coli QZ1111 could accumulate succinate with a concentration of 26.4 g/L under aerobic conditions. The byproduct acetate was only 2.3 g/L. The production ratio of succinate and acetate reached 11.5:1.

Abstract:Spiramycin and midecamycin are 16-membered macrolide antibiotics with very similar chemical structures. Spiramycin has three components, namely spiramycin I, II and III. Spiramycin II and III are, respectively, the O-acetyl and propionyl derivatives at C3-hydroxyl group of spiramycin I. Midecamycin has four components, and the C3-hydroxyl group of midecamycin is all O-propionylated. The enzyme adding acyl group(s) at the C3-hydroxyl group during the biosynthesis of spiramycin and midecamycin is 3-O-acyltransferase. The 3-O-acyltransferases for spiramycin and midecamycin are also very similar, and presume to function when exchanged. To explore whether the 3-O-acyltransferase for midecamycin biosynthesis hold still the character of selective and efficient propionylation for spiramycin I at its C3-hydroxyl group, we inserted mdmB, the 3-O-acyltransferase gene from Streptomyces mycarofaciens ATCC 21454 for midecamycin biosynthesis, into a mutant strain of S. spiramyceticus F21, in which the 3-O-acyltransferase gene for spiramycin biosynthesis, sspA, was deleted; and the mdmB was integrated exactly into the chromosomal site where the sspA was deleted. We name this “hybrid” strain as SP-mdmB. HPLC analysis of the spiramycin produced by SP-mdmB showed that spiramycin I was still the major component, although the relative proportions of both spiramycin II and III increased significantly. We thus conclude that MdmB from Streptomyces mycarofaciens ATCC 21454 for midecamyicn biosynthesis do not hold the character of selective and efficient propionylation for spiramycin I within S. spiramyceticus F21, and this character is possibly limited in Streptomyces mycarofaciens ATCC 21454 for midecamycin biosynthesis.

Abstract:We developed the recombinant green fluorescent protein gene yeast cell to screen estrogenic compounds based on two episomal vectors. In the expression vector the expression of human estrogen receptor a(hERa) was driven by 3-glyceraldehyde- phosphate dehydrogenase (GPD) promoter; in the reporter vector the expression of the yeast enhanced green fluorescent protein (yEGFP) gene was under the control of the estrogen response element (ERE). The vectors were transformed into yeast cell (W303-1A) to construct GFP recombinant yeast cell. Incubation of the yeast cell with various concentrations of the estrogenic compounds led to expression of the reporter gene product GFP in a dose dependent manner. Compared to other yeast bioassays, the yeast cell for environmental estrogen bioassay based on yEGFP reporter gene did not need cell wall disruption or the addition of a substrate or reagent. This yEGFP assay was performed completely in 96 well plates. So this test system can be used as a rapid and high throughput system for screening estrogenic chemical products, which has the characteristics of the sensitivity, reproducibility and cheapness.

Abstract:Hepatitis B virus (HBV) infection can cause the severe threat to the health of the people around the world. It depends upon the development of efficient diagnostic reagent and vaccine to prevent the prevalence of HB. In this study, we constructed the high expression recombinant Pichia pastoris and performed the screening tests in shake flasks to obtain the optimal values of several key fermentation parameters. Based on their effects on the growth and expression level of recombinant strains, FBS was the optimal industrial medium. The optimal values for the dissolve oxygen (DO), the final concentrations of methanol and the pH values were 50 mL, 1% (V/V) and 5.4–6.0, respectively. The optimal values of the parameters simulated in shake flasks were successfully scaled up to 10 L bioreactors to achieve high-throughput production: 310 OD600 in biomass and 27 mg/L in recombinant HBsAg. The expressed recombinant HBsAg in P. Pastoris was confirmed by SDS-PAGE and Western blotting. Electron microscopy examination showed that the purified protein could be self-assembled to 22 nm virus-like particles. The results provided a basis for industrial scale-up production of diagnostic reagent and vaccine of next generation against HB.

Abstract:Most studies related to determining the expression profile of genes and specific promoters used histochemical localization of the reporter gene, gusA. While the histochemical method for visualizing gusA expression suffers from several limitations in the determination of gene expression and location, especially in the tissues with high background acitivty. To solve this problem, a transient expession vector pBI221-RFP/GFP, was constructed using GFP and RFP as double fluorescent marker genes. This vector used CaMV 35S promoter to drive GFP and determine the transforming efficiency. It analyzed expression profile of the target gene and promoter through the RFP activities of the tranformed tissues. Through the specific promoter AGPL1 from watermelon and E8 promoter from tomato, it is resistible to use this vector to study the expression patterns of promoters. Results indicated that the pBI221-RFP/GFP is a very efficient transient expression vector that can be verify the functions of the genes and promoters.

Abstract:Damage effects of water sorption on mechanical properties of the hydroxyapatite particle reinforced Bis-GMA/TEGDMA copolymer (HA/Bis-GMA/TEGDMA) have been predicted using 3-D finite cell models. Three different cell models were used to determine the influence of varying particle contents, interphase strength and moisture concentration on the debonding damage. The stress distribution pattern has been examined and the stress transfer mode has been clarified. The Young’s modulus and fracture strength of the Bis-GMA/TEGDMA composite were also predicted using the model with and without consideration of the damage. The former results with consideration of the debonding damage are in good agreement with existing literature experimental data. The shielding effect of our proposed model and an alternative approach were discussed. The FCC cell model has also been extended to predict the critical load for the damaged and the undamaged composite subject to the 3-point flexural test.

Abstract:Following Escherichia coli lysis with alkali, cetyltrimethylammonium bromide (CTAB) was directly titrated into the supernatant. An easy and feasible technology for plasmid purification was established with the optimized proportion between the quantity of CTAB and plasmid, combined with the specific solution for DNA release and TritonX-114 for endotoxin removal. Quality detection showed that the purified plasmid was free of contamination of host RNA. The host bacterial genomic DNA, endotoxin and bacterial protein were less than 10 μg, 50 EU and 10 μg per mg plasmids, respectively. The ratio of OD260/OD280 was between 1.75-1.85. Eighty percent of the prepared plasmids were presented in the supercoiled form. The plasmid purified with this technology can satisfy all criteria stipulated by FDA. The main advantages of the technology include the avoidance of animal-derived enzymes such as ribonucleases A, Proteinase K and toxic reagents like chloroform and phenol. In addition, the technology has low cost and no pollution.

Abstract:There is no information

Abstract:D-amino acid oxidase from Trigonopsis variabilis (TvDAAO) was overproduced in E. coli cells and properties of the recombinant enzyme were studied. Single point mutants of the enzyme with 2.4-fold higher thermal stability or changed spectra of substrate specificity compared to wild-type enzyme were prepared. It was shown that mutant TvDAAO has higher catalytic efficiency in cephalosporin C oxidation in comparison with wild-type enzyme. One mutant of recombinant TvDAAO was crystallized and its structure was solved with resolution 2.8 ?.

Abstract:Acidic proteins are generally thought to control mineral formation and growth. Thus, characterization of acidic proteins in the insoluble organic matrix is an important first step toward linking function to individual proteins in alcyonarian coral. Analysis of proteinaceous components in the soluble and insoluble matrix fractions of Sinularia polydactyla indicates that aspartic acid composes about 61% of the insoluble and 29% of the soluble matrix fractions. Using an in vitro assay, we show that matrix proteins induced formation of amorphous CaCO3 precipitates prior to their transformation into the calcitic crystalline form. The crystalline form of CaCO3 in the sclerites was also identified by X-ray diffraction, revealing calcitic polymorphisms with a strong (104) reflection. The structure of alcyonarian organic matrices containing aspartate-rich proteins and polysaccharides was assessed by Fourier transform infrared spectroscopy (FT-IR). Calcium-binding analysis of components in the insoluble matrix fraction indicated that a protein of 109 kDa can bind Ca2+, which is important for sclerite formation. An assay for carbonic anhydrase (CA) enzyme, which is thought to play an important role in the process of bio-calcification revealed novel activity. These results strongly suggest that the aspartic acid–rich proteins within the insoluble matrix of alcyonarians play a key role in biomineralization regulation.

Abstract:We expressed an L-amino acid deaminase (Pma) from Proteus mirabilis in Escherichia coli and characterized the kinetics of phenylpyruvic acid production. P. mirabilis Pma was well expressed in E. coli in an active state and was found to be associated with membranes. The association of Pma with cellular membranes is likely to be necessary for its enzymatic activity.

Abstract:We developed a new method that yields biologically potent therapeutic proteins such as interferon a2b (IFNa2) and human growth hormone (hGH) in high yields and with increased serum half-lives when expressed as arabinogalactan-protein (AGP) chimeras in cultured tobacco cells. The chimeric glycoproteins typically gave 500–1800 fold greater secreted yields than their non-glycosylated controls. Importantly, the chimeric glycoproteins showed an increased in vivo serum half-life of up to 13 fold while their biological activities remain similar to native IFNα2 and hGH.

Abstract:1-aminocyclopropane-1-carboxylate (ACC) synthase is the rate-limiting enzyme in ethylene biosynthesis pathway in plants, which catalyzes the conversion from S-adenosylmethionine (SAM) to 1-aminocyclopropane-1-carboxylate (ACC). The ACC synthase (ACS) is encoded by a multiple gene family and its expression is differentially regulated by a complicated network composed of biotic and abiotic signals responding to both internal and external stimuli under the translational and posttranslational levels. However, its expression regulatory mechanism now still remains incompletely clear. Recent years, several researches showed that the Natural Antisense Transcripts (NATs) played a critical role in gene expression regulation in eukaryotic organism, which attracted increasing attentions. Recently, a putative ACC synthase cis–NAT was predicted in Arabidopisis by the genomic-wide scanning technique. In this study, a cis-natural antisense transcript (cis-NATs) of ACC synthase gene, named ASACS2, was identified from six northeastern soybean cultivars of China by RT-PCR and bidirectional sequencing. Further investigation by Real-time RT-PCR showed that during the vegetative growth stage ASACS2 co-existed with its higher-abundant sense counterparts, SACS2. Interestingly, Real-time RT-PCR also showed that the ratio between ASACS2 and SACS2 remain constant in each cultivar, but varied among six cultivars, suggesting the cultivar specificity. This study indicates that ASACS2 might be a potential regulatory actor in ethylene metabolism pathway. The further knowledge of this actor would facilitate us to better understand the gene regulation machinery in plants and the application in transgenic research.

Abstract:The archaeocyte-dominant cell population (ADCP) primmorphs is a new sponge cell culture system and possible to develop into a continuous sponge cell line. However, the archeaocytes’ division, differentiation and development as well as the cell interactions have not yet been clear. Cell movement (cell locomotion) is the base of other cell interactions. We use real-time video cinemicrography technique to document the cell locomotion continuously up to 90 days in the ADCP cultures of marine sponge Hymeniacidon perlevis. Locomotion of three typical cell types, archeaocytes for mesohyl cells, pinacocytes for surface cells and spicule-associated cells, were monitored and analyzed during the culture process including the inoculation of dissociated cells and the formation stage of functional primmorphs. We observed the unique particle transfer process between archaeocytes, the fluctuation spreading over cells and the production of silica spicules and the active transfer of spicules by sponge cells. A tentative model of material transfer and the coordinated locomotion of sponge cells were proposed.

Abstract:The integration of omics data in metabolic flux analysis is critical to explore the cellular mechanisms in the post-genomic era. Enzyme activity data can be integrated into elementary modes (EMs) by Enzyme Control Flux (ECF) for the prediction of flux distributions of mutants. A new ECF algorithm, named as ECFMEP, is proposed for a moderate scale of metabolic networks. To efficiently estimate a large number of EM coefficients (EMCs), the Lagrange multiplier is applied to the optimization problem under maximum entropy principle (MEP), thereby remarkably reducing the number of the variables to be explored. ECFMEP is employed to predict the flux distribution of four mutants of Escherichia coli under anaerobic conditions. These cells consist of 102 reactions and 28,555 EMs. We demonstrate that ECFMEP effectively makes use of enzyme activity data for enhanced prediction accuracy in comparison with that by Flux Balance Analysis (FBA) and Minimization of Metabolic Adjustment (MOMA).

Abstract:The primary human hypertrophic scar and normal skin fibroblasts were successfully established and identified by HSP47 and FSP markers. On the other hand, different pattern of protein expressions were found in cultured fibroblasts from hypertrophic scar and normal fibroblasts skin cells after treatment with the chitosan derivatives sheet. The CTCF protein was up-regulated in fibroblasts hypertrophic scar upon treatment with chitosan derivatives. In contrast, the amount of CTCF protein was found unchanged in normal skin fibroblasts both treated and untreated. The YB-1 protein was expressed almost similarly in normal and hypertrophic scar when treated with chitosan but the expression differed when untreated. The c-myc and p53 proteins were expressed in fibroblasts hypertrophic scar followed by up-regulation after chitosan derivatives treatment. The c-myc and p53 expressions were not detected in normal fibroblasts neither untreated nor treated. The CTCF, YB-1, c-myc and p53 proteins behaved in different manners in human hypertrophic scar and normal fibroblasts skin cells. The novel chitosan derivatives sheet in this study may play roles in the control of cell growth and proliferation of human hypertrophic scar and normal fibroblasts skin cells. The mechanisms underlying expression of these protein factors remain unclear and further studies are still undergoing in our laboratory.

Abstract:During an investigation of plant cell cultures which may be useful for the treatment of renal disorders, we established a well-growing E-4 callus culture of Eritrichium sericeum that produced high amounts of caffeic acid metabolites, (–)-rabdosiin (1.8% dry wt) and rosmarinic acid (4.6% dry wt). Elicitation of the calli induced an increase in (–)-rabdosiin production by as much as 4.1% dry wt. Oral administration of E-4 callus biomass to rats with induced Masugi-nephritis caused an increase of diuresis, lowered creatinine excretion and proteinuria levels, compared with Masugi-nephritis untreated rats. While all of the Masugi-nephritis untreated rats began to ache, near a quarter of the E-4 treated rats remained in good health. This result indicates that the E-4 culture has a potential to alleviate symptoms associated with nephritis. A mechanism by which production of caffeic acid metabolites could be activated in the calli was studied using a high polyphenol-producing cell cultures transformed with the rolC gene. We established that the increase of caffeic acid metabolites production in rolC-transgenic E. sericeum calli positively correlated with high expression of the CYP98A3 gene, a key gene for rosmarinic acid biosynthesis.

Abstract:A major concern in developing protein-based biopharmaceuticals is protein instability. A strategy with the use of reducing agent pretreatment to improve protein stability was developed for recombinant hCD83ext (i.e. the extracellular domain of human CD83) with a potential therapeutic activity. Under physiological conditions, the therapeutic product tended to denature, form aggregates and precipitates, and eventually degrade. The reducing agent pretreatment was demonstrated to be effective in improving the protein stability.