Hui Ruan

Optimization of ultrasonic-assisted extraction (UAE) of betulin from white birch bark using response surface methodology.

Betulin is an abundant naturally occurring triterpene, which makes it a potentially important raw material for a precursor of biologically active compounds. The objective of the current study was to determine the optimum UAE conditions for betulin from B. papyfera bark. The optimum conditions were evaluated with fractional factorial design and optimized using response surface methodology. High yields of betulin were observed from white birch bark by UAE technology. The solvent concentration and the ratio of material to solvent were the most significant parameters on betulin extraction as evaluated through FFD. The extraction conditions were further investigated with central composite design. The fitted second-order model revealed that the optimal conditions consisted of 98% ethonal concentration, 1:42 the ratio of white birch bark to solvent, extraction temperature 50 degrees C, ultrasonic frequency 5kHz and extraction time 3h. Under the optimized condition, the maximum productivity of betulin predicted is 23.17%. The extraction productivity and purity of betulin under the optimized extraction conditions were great higher than that of the non-optimized condition. The present study demonstrates that ultrasound is a great efficiency tool for the fast extraction of betulin from white birch bark.

Isolation and characterisation of an oxygen, acid and bile resistant Bifidobacterium animalis subsp. lactis Qq08

BACKGROUND Currently, bifidobacteria are recognised as one of the most important bacteria used as probiotics, which promote human health. However, their commercial application has been limited by their anaerobic nature. The purpose of this study was to select an oxygen, acid and bile resistant strain of bifidobacterium for use as a new probiotic. RESULTS A total of 10 strains of bifidobacteria from different sources were analysed for their relative bacterial growth ratio (RBGR) in different oxygen concentrations. Three strains with high RBGR values were selected and their survival rates in acid environment and bile salt conditions were investigated in vitro. One strain showed high tolerance to low pH, giving a survival rate of 84% at pH 2 after 4 h incubation, and high tolerance to bile, more than 90% after 4.5 h incubation at 0.01 g mL(-1) bile concentration. This strain was identified as Bifidobacterium animalis subsp. lactis strain Qq08 based on polyphasic taxonomy approaches, such as phenotype analysis and 16S rRNA and 16S to 23S internally transcribed spacer sequence analyses. CONCLUSION We isolated an aerotolerant bifidobacterium and identified it as Bifidobacterium animalis subsp. lactis Qq08. This strain has characteristics more favourable than the commercial probiotic strain, Bifidobacterium lactis Bb12.

Studies on optimization of nitrogen sources for astaxanthin production by Phaffia rhodozyma

Fermentation of Phaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using (NH4)2SO4, KNO3 and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L (NH4)2SO4, 0.49 g/L KNO3 and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.

Production of conjugated linoleic acids by Lactobacillus plantarum strains isolated from naturally fermented Chinese pickles

Naturally fermented pickles harbour many lactic acid bacteria (LAB). Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, lp15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 μg/ml of linoleic acid (LA). Compared to other strains, L. plantarum strain lp15 showed the highest tolerance upon increased levels of LA in the medium, i.e., up to 600 μg/ml. This strain converted about 25% of LA into CLA isomers [predominantly cis-9, trans-11 CLA (9-CLA) and trans-10, cis-12 CLA (10-CLA)], of which 75% was 9-CLA. Interestingly, though the conversion rate of LA into CLA by lp15 remained stable between 100 to 600 μg/ml LA levels in the medium, it dropped sharply at 1000 μg/ml. Taken together, the lp15 strain displayed relatively high LA tolerance with higher conversion rate, which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08

Biotransformation of phytosterol (PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper. The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology. Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M. neoaurum ZJUVN-08 strain. Results showed that molar ratio of hydroxypropyl-β-cyclodextrin (HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production. By analyzing the statistical model of three-dimensional surface plot, the optimal process conditions were observed at 0.1 g/L inducer, pH 7.0, molar ratio of HP-β-CD to PS 1.92:1, 8.98 g/L PS, and at 120 h of incubation time. Under these conditions, the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized (5.99 g/L), while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized (84.03%). The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results. It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process.

Investigation on Culturable Microflora in Tibetan Kefir Grains from Different Areas of China

UNLABELLED Four samples of Tibetan kefir grains (TK-ZJUJ 01-04) from Tibet and surrounding areas were investigated via phenotypic and genotypic methods to compare and analyze the diversity of culturable microflora among different origins. As a result, 4 genera of microorganisms from TK-ZJUJ01: Bacillus subtilis (2.9 × 10(7) cfu/mL), Lactococcus lactis (8.2 × 10(7) cfu/mL), Kluyveromyces marxianus (3.0 × 10(6) cfu/mL), Saccharomyces cerevisiae (9.0 × 10(6) cfu/mL); 4 genera from TK-ZJUJ02: Lactobacillus kefiri (1.0 × 10(8) cfu/mL), Pichia kudriavzevii (5.0 × 10(6) cfu/mL), K. marxianus (1.9 × 10(7) cfu/mL), Kazachstania unispora (6.2 × 10(7) cfu/mL); 6 genera from TK-ZJUJ03: Leuconostoc lactis (4.6 × 10(7) cfu/mL), L. lactis (3.0 × 10(7) cfu/mL), Lactobacillus plantarum (3.0 × 10(7) cfu/mL), K. unispora (3.0 × 10(6) cfu/mL), K. marxianus (2.0 × 10(6) cfu/mL), (1.7 × 10(7) cfu/mL); and 4 genera from TK-ZJUJ04: L. plantarum (1.8 × 10(7) cfu/mL), Acetobacter fabarum (5.0 × 10(6) cfu/mL), K. unispora (6.2 × 10(7) cfu/mL), Pichia guilliermondii (6.2 × 10(7) cfu/mL) were identified. Yeasts like P. kudriavzevii and P. guilliermondii isolated in this study were the first time reported in Tibetan kefir grains. For TK-ZJUJ 01-03, lactic acid bacteria were the major microorganisms, which accounted for more than 50% of all the microbial population, while for TK-ZJUJ04, the largest microbial group was yeasts which accounted for more than 50%. In a word, study of diversity and composition of microflora provided us theoretical foundation for further investigation and application of Tibetan kefir grains. PRACTICAL APPLICATION This is the basic research in order to develop and industrialize a new kind of yogurt starter which is naturally formed microbiota with both lactic acid bacteria and yeasts in it.

Hydrolyzates of silkworm pupae (Bombyx mori) protein is a new source of angiotensin I-converting enzyme inhibitory peptides (ACEIP).

Silkworm pupae protein is a good source of high quality protein. The hydrolyzates of silkworm pupae protein catalyzed by neutrase, pepsin, acidic protease (Asperqiius usamii NO. 537), flavourzyme, alcalase, and trypsin with inhibitory activity on angiotensin I-converting enzyme (ACE) were identified by HPLC. The hydrolyzates catalyzed by acidic protease exerted the highest inhibitory activity on ACE. The hydrolyzing conditions were optimized by one-factor, factional factorial (FFD), and center composite (CCD) design methods, and response surface methodology (RSM). Statistical analyses showed that regression of the second-order model equation is suitable to describe ACE inhibitory bioactivity. The predicted inhibitory activity of hydrolyzates on ACE was 73.5 % at a concentration of 2.0 mg/ml. Optimized RSM technique decreased IC(50) of hydrolyzates inhibiting ACE to 1.4 mg/mL from 2.5 mg/ml. The molecular weight of the components of the hydrolyzates with inhibitory activity on ACE varied from less than 500 to about 1000 Da by ultra-filter analysis. These studies suggest that hydrolyzates of silkworm protein contain ACE inhibitory activity that could form a potential source of ACE inhibitor drugs.

Response surface methodology for optimizing the fermentation medium of alpha-galactosidase in solid-state fermentation.

Aims:  Alpha‐galactosidase is applied in food and feed industries for hydrolysing raffinose series oligosaccharides (RO) that are the factors primarily responsible for flatulence upon ingestion of soybean‐derived products. The objective of the current work was to develop an optimal culture medium for the production of alpha‐galactosidase in solid‐state fermentation (SSF) by a mutant strain Aspergillus foetidus.

Studies on a kinetic model for butyric acid bioproduction by Clostridium butyricum

Aims:  This paper discusses the establishment of a kinetic model for cell growth, butyric acid production and substrate consumption of Clostridium butyricum ZJUCB in batch cultivation.

Mechanism of Free Zn(2+) Enhancing Inhibitory Effects of EGCG on the Growth of PC-3 Cells: Interactions with Mitochondria

Green tea and its major constituent epigallocatechin gallate (EGCG) are known for their chemopreventive effects including those against prostate cancer, which could be mediated by metal ions. Zn2+ is an essential trace element that is required for human health and plays an important role in the normal function of the prostate gland. In the present study, the effect of EGCG on cell membrane and mitochondria of PC-3 (prostate carcinoma) cells in the presence and absence of Zn2+ was studied. These studies revealed that EGCG, Zn2+, or EGCG + Zn2+ affected the morphology of PC-3 cells and induced apoptosis in PC-3 cells. It was observed that effects of treatment with EGCG, Zn2+, or EGCG + Zn2+on mitochondria showed EGCG + Zn2+ > Zn2+ > EGCG, including cytochrome C release from the intermembrane space into the cytosol, inhibited the synthesis of ATP, loss of mitochondrial membrane potential, and activation of caspase-9. However, the order of effect on depressing membrane fluidity of PC-3 cells was EGCG > EGCG + Zn2+ > Zn2+. In summary, these findings suggest that EGCG, Zn2+, and EGCG + Zn2+ induce necrosis or apoptosis of PC-3 cells through mitochondria-mediated apoptotic pathway and free Zn2+-enhanced effects of EGCG on PC-3 cells due to its interactions with mitochondria.