Guowei Shu

Effect Of Five Proteases Including Alcalase, Flavourzyme, Papain, Proteinase K And Trypsin On Antioxidative Activities Of Casein Hydrolysate From Goat Milk

Abstract Oxidation was related to the pathogenesis of human diseases. Adequate intake of antioxidant activity of food can reduce the levels of free radicals, prevent lipid peroxidation, and help the body against diseases. In the paper, casein from goat milk was hydrolyzed by five commercial proteases, namely, Alcalase, flavourzyme, papain, proteinase K and trypsin. The antioxidant activities of casein hydrolysates were assessed by evaluating hydrolysis degree, DPPH radical-scavenging activity, metal-chelating activity and superoxide radical scavenging activity. The results showed as follows: the DH of proteinase K, Alcalase, and trypsin were higher significantly than those of papain and flavourzyme. The Fe2+-chelation activity and superoxide radical scavenging activity of casein hydrolysates from goat milk by Alcalase was higher than the others, the DPPH scavenging activities of casein hydrolysates by Alcalase and papain was higher than the others and the DPPH scavenging activities by Alcalase and papain had no significant diffierence (p<0.05), so the optimal proteinase for hydrolysis casein from goat milk to produce antioxidant peptide was Alcalase.

Effect of incubation time, inoculum size, temperature, pasteurization time, goat milk powder and whey powder on ace inhibitory activity in fermented milk by L. Plantarum LP69

BACKGROUNDnAngiotensin I converting enzyme (ACE) plays an important physiological role in regulating hypertension. Lactic acid bacteria are known to produce ACE inhibitory peptides which can lower hypertension during fermentation.nnnMETHODSnThe effect of incubation time (0~36 h), inoculum size (3, 4, 5, 6 and 7%, v/v), temperature (25, 30, 35, 40 and 45°C), sterilization time (5, 10, 15, 20 and 25 min), concentration of goat milk powder (8, 10, 12, 14 and 16%, w/v) and whey powder (0.5, 0.6, 0.7, 0.8 and 0.9%, w/v) on ACE inhibitory peptides fermented from goat milk by Lactobacillus plantarum LP69 was investigated using single factor experiment.nnnRESULTSnThe optimal incubation time, inoculum size, temperature, pasteurization time, goat milk powder and whey powder in fermented milk by L. plantarum LP69 was 14 h, 3.0%, 35°C, 20 min, 14% and 0.70% for ACE inhibitory activity and 22 h, 3.0%, 40°C, 25 min, 16% and 0.60% for viable cell counts, respectively.nnnCONCLUSIONSnThe incubation time, inoculum size, temperature, pasteurization time, goat milk powder and whey powder had a significant influence on ACE inhibitory activity in fermented milk by Lactobacillus plantarum LP69, the results are beneficial for further screening of main factors by using fractional factorial designs.

Characterization of Goat Milk Hydrolyzed by Cell Envelope Proteinases from Lactobacillus plantarum LP69: Proteolytic System Optimization, Bioactivity, and Storage Stability Evaluation

Despite the widespread application of lactic acid bacterium in dairy production through its contribution to acidification, development of sensorial properties, and health-promoting effects, relatively little information is available on the cell envelope proteinases (CEPs) of Lactobacillus plantarum, especially on the proteolytic system and the production of bioactivity peptides. In this study, CEPs from a novel L. plantarum LP69 were involved in goat milk hydrolysis and generated a product with high activity that showed a degree of hydrolysis of 15.68 ± 0.74%, Angiotensin I-Converting Enzyme (ACE)-inhibitory rate of 83.25 ± 1.05%, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging rate of 64.91 ± 1.27%, and hydroxyl radical scavenging rate of 89.17 ± 1.13%. The optimized hydrolysis conditions were time of 4.5 h, temperature of 41 °C, initial pH of 8.5, and enzyme to substrate ratio (E/S) of 12% (w/w) by orthogonal experiments. Application of a stabilizer greatly promoted milk stability. A well-designed stabilizer consists of 0.05% carrageenan, 0.15% gellan gum, and 0.15% sucrose esters, which significantly raised the milk stability coefficient, R, from 70.67% to 98.57%. The storage stability of milk was evaluated during 84 days at room temperature or 4 °C. Our study depicts the contribution of CEPs from L. plantarum LP69 in goat milk, exploring a new way for the development of a functional milk product.

Microencapsulation of Lactobacillus Acidophilus by Xanthan-Chitosan and Its Stability in Yoghurt

Microencapsulations of Lactobacillus acidophilus in xanthan-chitosan (XC) and xanthan-chitosan-xanthan (XCX) polyelectrolyte complex (PEC) gels were prepared in this study. The process of encapsulation was optimized with the aid of response surface methodology (RSM). The optimum condition was chitosan of 0.68%, xanthan of 0.76%, xanthan-L. acidophilus mixture (XLM)/chitosan of 1:2.56 corresponding to a high viable count (1.31 ± 0.14) × 1010 CFU·g−1, and encapsulation yield 86 ± 0.99%, respectively. Additionally, the application of a new encapsulation system (XC and XCX) in yoghurt achieved great success in bacterial survival during the storage of 21 d at 4 °C and 25 °C, respectively. Specially, pH and acidity in yogurt were significantly influenced by the new encapsulation system in comparison to free suspension during 21 d storage. Our study provided a potential encapsulation system for probiotic application in dairy product which paving a new way for functional food development.

Optimization of Goat Milk with ACE Inhibitory Peptides Fermented by Lactobacillus bulgaricus LB6 Using Response Surface Methodology

In the present study, the incubation conditions of goat milk fermented by Lactobacillus bulgaricus LB6 were optimized to increase the angiotensin converting enzyme (ACE, EC 3.4.15.1) inhibitory activity by Box–Behnken design of response surface methodology. Incubation temperature, whey powder, and calcium lactate had significant effects on ACE inhibition rate and viable counts of LB6 during incubation. The results showed that optimal conditions of fermentation were found to be 37.05 °C, 0.8% (w/w) whey powder and 0.50% (w/w) calcium lactate. ACE inhibition rate increased significantly from 71.04 ± 0.37% to 83.31 ± 0.45% and the viable counts of Lactobacillus bulgaricus LB6 reached to 8.03 × 107 cfu·mL−1 under the optimal conditions, which approached the predicted values 83.25% and 8.04 × 107 cfu·mL−1. The optimal fermentation conditions can be a good reference for preparing ACE inhibitory peptides from goat milk.

Microencapsulation of Bifidobacterium bifidum BB01 by xanthan–chitosan: preparation and its stability in pure milk

Abstract Xanthan–chitosan (XC) and xanthan–chitosan–xanthan (XCX) were employed for microencapsulation of Bifidobacterium bifidum BB01 using extrusion technique. To optimize the process of B. bifidum BB01 microcapsules based on XC hydrogels, response surface methodology was employed to obtain the best possible combination of chitosan concentration, xanthan-B. bifidum BB01 mixture (XBM)/chitosan, stirring time for the maximum viable count and encapsulation yield. The optimum conditions were: chitosan concentration of 0.84u2009g·mL−1, XBM/chitosan of 1:9.0, stirring time of 60u2009min with high viable count and encapsulation yield of 1.52u2009±u20090.15u2009×u20091010 CFU·g−1, 90u2009±u20090.65%, respectively. In addition, the effective encapsulation system (XC and XCX) resulted in improvement in survival of B. bifidum BB01 compared to non-encapsulated cells during 3u2009weeks storage at 4 and 25u2009°C in pure milk.

Enzymolysis Technology Optimization for Production of Antioxidant Peptides from Goat Milk Casein

Abstract Antioxidant peptides can inhibit lipid peroxidation and scavenging free radicals, maintain the balance of free radicals, and against a variety of diseases. Response surface methodology was used to optimize process conditions for producing antioxidative peptides from goat’s milk casein hydrolysate with Alcalase. The results suggested that the optimal process parameters were: temperature at 62.5°C, pH 8.9, E/S ration at 2.5%, substrate concentration at 4.4% and hydrolysis time was 173min). Metal-chelating effect, superoxide anion radical scavenging activity and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity were shown to be 87.21±0.88%, 49.18±1.42% and 69.07±1.26% respectively under the optimal condition. The actual and predicated value were closely which indicated the optimized data fit well to model and the optimized parameters are reliable.

Optimization of nitrogen source for Bifidobacterium bifidum using response surface methodology

Abstract In order to improve the viable counts of Bifidobacterium bifidum BB01 in the liquid medium, the Central Composite Design (CCD) was used to optimize the nitrogen source in the medium of B. bifidum BB01. The results showed that the nitrogen source composition of B. bifidum BB01 was: peptone 0.9%, yeast extracts 0.3%, beef paste 0.7%. Under the optimal conditions, the viable counts of B. bifidum BB01 reached (2.49±0.06)×109CFU/mL after cultured at 18h, which was 42.97% higher than MRS (lactose), and 12.85% higher than the optimized MRS medium (carbon source and prebiotics were optimized). Therefore, the CCD used in this study is workable for promoting the growth of B. bifidum BB01.

Optimization of the medium for Lactobacillus acidophilus by Plackett-Burman and steepest ascent experiment.

BACKGROUNDnLactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment.nnnMETHODSnThe ideal carbon source was screened among glucose, maltose, lactose and whey powder, and the ideal nitrogen source was screened among casein hydrolysate, peptone, yeast extract powder, fish meal, carbamide, ammonium sulfate and sodium nitrate by single factor experiment. Plackett-Burman and steepest ascent experiment were applied to screen the main effective factors of Lactobacillus acidophilus among peptone, beef extract, yeast extract powder, glucose, K2HPO4, C6H14O7N2, CH3COONa, MgSO4 and Tween-80. Result. The results indicated that glucose (p = 0.01510) as negative factor and K2HPO4 (p = 0.02017) as positive effect were the significant growth factors of Lactobacillus acidophilus, CH3COONa (p = 0.09273) as positive effect was an important factor, and the optimized medium was as follows: glucose - 21 g/L, K2HPO4 - 3.5 g/L, CH3COONa - 6.5 g/L, peptone - 10 g/L, beef extract - 8 g/L, yeast extract pow.nnnRESULTSnnd. Lactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment. Material and methods. The ideal carbon source was screened among glucose, maltose, lactose and whey powder, and the ideal nitrogen source was screened among casein hydrolysate, peptone, yeast extract powder, fish meal, carbamide, ammonium sulfate and sodium nitrate by single factor experiment. Plackett-Burman and steepest ascent experiment were applied to screen the main effective factors of Lactobacillus acidophilus among peptone, beef extract, yeast extract powder, glucose, K2HPO4, C6H14O7N2, CH3COONa, MgSO4 and Tween-80. Result. The results indicated that glucose (p = 0.01510) as negative factor and K2HPO4 (p = 0.02017) as positive effect were the significant growth factors of Lactobacillus acidophilus, CH3COONa (p = 0.09273) as positive effect was an important factor, and the optimized medium was as follows: glucose - 21 g/L, K2HPO4 - 3.5 g/L, CH3COONa - 6.5 g/L, peptone - 10 g/L, beef extract - 8 g/L, yeast extract powder - 8 g/L, C6H14O7N2 - 2 g/L, MgSO4 - 0.2 g/L and Tween-80 - 1 mL/L when the maximum viable count could achieve 2.72·109 cfu/mL.nnnCONCLUSIONSnThe experimental model is reliable and the experimental results are of good stability. Variance analysis is performed to determine the adequacy and significance of the linear model. Thus, Plackett-Burman and steepest ascent experiment improve the veracity of optimization the medium for Lactobacillus acidophilus compared with the previous research.

Screening of probiotic goat milk tablets using Plackett-Burman design.

Background. Probiotics defi ned as additional microorganisms were added to goat milk powder, which not only improves the intestinal fl ora balance but also promotes human and animal health. The objectives of this study were to improve and guarantee high probiotics viable count and accordance with consumer’s acceptance. Material and methods. The reading selected the number of colony between 30 and 300, then calculated the viable count per gram of goat milk tablet (cfu/g). The items of sensory evaluation included: appearance, fl avour, colour, texture and taste. The score test was composed of 5 trained assessors, scored combination of different formulations (full marks of 100 points) and recorded the results. Results. Analysis of the results showed that sucrose, inulin and mannitol were selected as the main effective parameters on both viable count and sensory evaluation. Furthermore optimization of the formulation of probiotic goat milk tablets was to maximise the probiotics viable count to achieve 9.5·10 8 cfu/g and its scores of sensory evaluation to get 94 points. Discussion. Future probiotics products will be combined with a variety of probiotics, which can display their respective advantages and characteristics. Thus the products will not only be in accordance with the requirements of human health and trend of social development, but also will quickly become a favorite among consumers.BACKGROUNDnProbiotics defined as additional microorganisms were added to goat milk powder, which not only improves the intestinal flora balance but also promotes human and animal health. The objectives of this study were to improve and guarantee high probiotics viable count and accordance with consumers acceptance.nnnMETHODSnThe reading selected the number of colony between 30 and 300, then calculated the viable count per gram of goat milk tablet (cfu/g). The items of sensory evaluation included: appearance, flavour, colour, texture and taste. The score test was composed of 5 trained assessors, scored combination of different formulations (full marks of 100 points) and recorded the results.nnnRESULTSnAnalysis of the results showed that sucrose, inulin and mannitol were selected as the main effective parameters on both viable count and sensory evaluation. Furthermore optimization of the formulation of probiotic goat milk tablets was to maximise the probiotics viable count to achieve 9.5·108 cfu/g and its scores of sensory evaluation to get 94 points.nnnCONCLUSIONSnFuture probiotics products will be combined with a variety of probiotics, which can display their respective advantages and characteristics. Thus the products will not only be in accordance with the requirements of human health and trend of social development, but also will quickly become a favorite among consumers.