Technology for Enzymolysis of Jellyfish Brain Protein by Bromelain

[Objective] This study aimed to investigate the optimal technology for en- zymolysis of jeUyfish brain protein by bromefain. [Method] Effects of enzymolysis temperature, the amount of added enzyme, solid/liquid ratio, enzymolysis time and pH on the enzymolysis of total protein were investigated through single factor ex- periments, and some technical parameters were optimized by orthogonal experi- ments. [Result] The results revealed that enzymolysis temperature of 55℃, the amount of added enzyme of 4 000 U/g, solid/liquid ratio of 1：4, the amount of added enzyme time for 5 h and pH 7 were the optimal combination of parameters for the enzymolysis; the average degree of hydrolysis obtained from the verification tests was 36.9%, indicating that the optimized results of the orthogonal experiment were credible. [Conclusion] This study provides the optimized conditions reliable theo- retical foundation and technical support for the exploitation and utilization of jellyfish resources as medicines or functional food in China.

Treatment of Enzymolysis Wastewater by Centrifugation- Coagulation- Fenton Reagent Oxidation- Adsorption Process

[ Objective ] The study aimed to discuss the optimal conditions for the treatment of enzymolysis wastewater by centrifugation - coagu- lation - Fenton reagent oxidation - adsorption process. [ Metbod] According to the water-quality characteristics of wastewater from a heparin so- dium production factory of Jiangsu Province, enzymolysis wastewater was segregated from intestinal lavage wastewater and treated through cen- trifugation- coagulation- Fenton reagent oxidation-adsorption process, and the optimal technical parameters were determined. E Resultl After enzymolysis wastewater was centrifuged at a speed of 4 000 rpm, 0.6 g/L CTS as the coagulant was added to the supematant. Hereafter, pH of the coagulated effluent was adjusted to 3, and then 1.5% （V/V） H2O2 was added to the coagulated effluent; a certain amount of ferrous sul- fate （n H2O2-.n FeSO4 . 7H2O =8：1 ） was added to the mixture; the reaction conducted for 30 min, and then solution pH was adjusted to about 9. Finally, the oxidized effluent flowed through a resin red until the adsorptive capacity reached 240 BV, and COD of the effluent water was lower than 100 mg/L, meeting the Grade-I standard of Comprehensive Discharge Standard of Sewage （GB8978-1996）. [Condusio] The research could provide a new process for the treatment of enzymolysis wastewater.

Oxidative depolymerization of lignin improved by enzymolysis pretreatment with laccase

A new lignin depolymerization approach for improving the yield of aromatic monomers（YAM） by enzymolysis pretreatment was investigated, in which lignin was pretreated with laccase followed by oxidative depolymerization. It was found that lignin depolymeirzation was enhanced significantly by enzymolysis. The oxidative depolymerization contributed to 21.37% of YAM after the enzymolysis pretreatment,whereas the conventional oxidative depolymerization only gave 14.10% of YAM. The addition of ethanol in enzymatic pretreatment process improved the efficiency of enzymolysis, which effectively improved the solubility of pretreated lignin and depolymerization degree（DD） of lignin. The enzymolysis pretreatment increased the content of syringyl（S） style aromatic monomers, which hindered the recondensation among polymerized products. As lignin has low solubility in acidic aqueous solution, ethanol was added into enzymolysis system to improve the efficiency. However, the enzymolysis of lignin should be carried out for a limited period of time to prevent the inactivation of laccase.

Study on the separation and mechanism of the enzymolysis of Dioscorea Zingiberensis

Based on a through study of the chemical compositions of Dioscorea Zingiberensis, a new separation technology is presented in this paper. This technology can effectively separate the starch and cellulose from Dioscorea Zingiberensis, thus the output of diosgenin is raised by 20%. Meanwhile using the suspension after separation, the synergy of enzymes, which are under the best condition for enzymolysis is studied, and an enzymolysis model, which uses two enzymes at the same time, is established. This model provides a theoretical basis for finding new enzyme resources. What＇s more orthogonal experiments show that by using this model, either the output of diosgenin can be increased by 50.58% or the amount of acid used can be reduced by 83% which means less pollution.

Formation Conditions of Aspergiilus oryzae Protoplast

[Objective] To explore the efficient way for the preparation of Aspergillus oryzae protoplast.[Method] The statistics of protoplast yield by the enzymolysis of mycelia with different culture time,different osmotic stabilizers and different concentrations of enzyme system was carried out in this study.[Result] The protoplast yield was higher when the mycelium was cultivated for 108 h,and the osmotic stabilizer was 0.8 mol/L of NaCl.With the enzymes system of 2 mg/ml of lysozyme,6 mg/ml of cellulose and 6 mg/ml of glusulase,the protoplast yield achieved its highest,which was up to 9.0×105 per ml.[Conclusion] The yield of protoplast was affected by the situation of mycelia themselves and external conditions.This study had provided a basis for the preparation of a large number of active protoplasts and the further researches on cell fusion.

High efficiency production of ginsenoside compound K by catalyzing ginsenoside Rb1 using snailase

The rare ginsenoside Compound K （C-K） is attracting more attention because of its good physiological activity and urgent need. There are many pathways to obtain ginsenoside C-K, including chemical and biological methods. Among these, the conversion of PPD-type ginsenosides by enzymatic hydrolysis is a trend due to its high efficiency and mild conditions. For effectively extracting from the other panaxadiol saponins, the conversion process for ginsenoside C-K was investigated using snailases in this study. The univariate experimental design and response surface methodology were used to determine the optimal hydrolysis conditions for the conversion of ginsenoside Rbl into ginsenoside C-K by snailases. The optimum conditions were as follows： pH 5,12, temperature 51 ℃, ratio of snailase/substrate 0.21, and reaction time 48 h. On the basis of these parameters, the addition of 1.0 mmol· L- 1 ferric ion was found to significantly improve the enzymolysis ofsnailases for the first time. With the above conditions, the maximum conversion rate reached 89.7%, suggesting that the process can obviously increase the yield of ginsenoside C-K. The bioassay tests indicated that the ginsenoside C-K showed anti-tumor activity in a series of tumor cell lines. Based on these results, we can conclude that the process of rare ginsenoside C- K production by enzymolysis with snailase is feasible, efficient, and suitable for the industrial production and application.

Arsenic Removal from Pinctada martensii Enzymatic Hydrolysate by Using Zr(Ⅳ)-Loaded Chelating Resin

The present study investigated the removal of inorganic arsenic from Pinctada martensii enzymatic hydrolysate through unmodified resin （D296） and Zr（IV）-loaded chelating resin （Zr-D401）. By loading Zr to macroporous chelating resin D401, the as exchange adsorption active sites are generated. This transforms D401 from a material that does not have the arsenic adsorption capacity into a material that has excellent arsenic exchange adsorption capacity. The static adsorption experiments were conducted to investigate the optimal removal condition for D296 and Zr-D401. The experimental results show that： the optimum condition for D296 is that T= 25℃, pH= 5, resin additive amount= 1 g （50 mL）-1, and contact time = 10 h, the corresponding arsenic removal rate being 65.7%, and protein loss being 2.33%; the optimum condition for Zr-D401 is that T=25 ℃, pH = 8, resin additive amount= 1 g （50 mL）-1, and contact time=10 h, the corresponding arsenic removal rate being 70.3%, and protein loss being 4.65%. These results show that both of the two resins are effective in arsenic removal for preserving useful substance. Our research provides scientific evidence and advances in the processing technology for heavy metal removal in shellfish.

Integration of Ru/C and base for reductive catalytic fractionation of triploid poplar

Lignin,which is the most recalcitrant component of lignocellulosic biomass,is also the most abundant renewable aromatic resource.Herein,reductive treatment of triploid poplar sawdust by the integration of catalytic Ru/C and a base,which afforded high yields of phenolic monomers from the lignin component and a solid carbohydrate pulp,is reported.The introduction of Cs_(2)CO_(3) led to the generation of C2 side‐chained phenols through the cleavage of C_(β)–O and C_(β)–C_(γ) bonds inβ–O–4 units in addition to C3 side‐chained phenols;the relationship between C2 and C3 was dependent on the base dosage.The reaction conditions,including base species,temperature,time,and H_(2) pressure,were optimized in terms of phenolic product distribution,delignification degree,and carbohydrate retention.The carbohydrate pulps generated from reductive catalytic fractionation in the presence of Cs_(2)CO_(3) were more amenable to enzymatic hydrolysis,indicating that this treatment of biomass constituted the fractionation of biomass components together with the breakdown of biomass recalcitrance.

A Comparative Study of the Difference Method and the Enzyme-Hydrolyzed Casein Method for Determining True Amino Acid Digestibilities and Endogenous Amino Acid Losses in Duck Feed

In this study, we examined the varia- tions between the difference method and the enzyme- hydrolyzed casein method for determining endogenous amino acid loss and the true amino acid digestibility in ducks fed normal protein-containing diets. These methods were compared to the nitrogen-free （N-free） diet method. The difference method was based on soy- bean meal as the only protein source, with the experi- mental diets containing crude protein levels at 15% and 20%. The enzyme-hydrolyzed casein method was based on enzyme-hydrolyzed casein meal as the pro- tein source, with the experimental diet containing a crude protein level of 17.5%. The N-free diet was prepared with starches and paper fibers. In each meth- od,64 Tianfu meat drakes （7-weeks-old） with an av- erage body weight of 2.77±0.16 kg were used and divided into four groups, and fed four different diets. Each group contained four replicates of four drakes and they were force fed trial diets according to the Sirbald method for detecting their apparent amino aciddigestibility, endogenous amino acid loss and true a- mino acid digestibility. The results demonstrated that using the difference, enzyme-hydrolyzed casein and N-free diet methods, endogenous amino acid losses were 0. 9946,1. 2243 and 0. 9297 mg/g dry matter in- take （ DMI）, respectively. The true amino acid digest- ibility measured by the difference method was 88.93 %±4.43 %. Using the enzyme-hydrolyzed ca- sein method with two dietary crude protein levels of 15% and 20%, the digestibility was 91.15%±4.33% and 91.97%±4. 16%, respectively, and by the N-free diet methods with two dietary crude protein levels of 15% and 20% ,it was 88.55%±4.29% and 88.82 %±4.61%, respectively. The results suggested that when the dietary protein level was 15% to 20 %, the true amino acid digestibility and endogenous ami- no acid loss as determined by the difference method was more accurate than the values determined by the enzyme-hydrolyzed casein method.

Optimization Conditions of Wheat Mesophyll Protoplast Isolation

The research object of this study is “ML7113” wheat leaf, which is used to isolate protoplast with enzyme hydrolysis method. Three main effectors—the concentration of mannitol, enzymolysis time and centrifugal force, affect the production and vitality of protoplast. While the production and vitality of wheat protoplasts were detected by the hemacytometer and the FDA staining respectively. Results showed that, with the increasing concentrations of mannitol during 0.2 M - 0.4 M, protoplast yield increases and when the concentration is 0.4 M, the protoplast vitality can be up to 95%;with the extension of enzymolysis time in 2 h to 8 h, protoplast yield reaches a maximum in 6 h, but its vitality achieves the maximum in 4 h;considering a combination of these two factors impacting on protoplast, we obtain the best time to digest for 4 h;meanwhile, with the increasing of the centrifugal force from 500 rpm - 2000 rpm, its comprehensive effect of protoplast vitality and yield is the highest when the centrifugal force is 1000 rpm for 2 min (replicated three times). So 0.4 M mannitol, 4 h enzymolysis time and 1000 rpm for 2 min centrifugal force are the best separation condition.

A Method of Purification of Bovine Colostrum sIgA

sIgA in bovine colostrum was purified by ultrafiltration and enzymolysis methods in this experiment, and the prepared substance was detected by Western Blot. The purity and yield were up to 73.6% and 65.2%, respectively. This convenient technique offers helpful exploration for production of bovine colostrum sIgA

A Two-step Biotechnological Process for Improving Nutrition Value of Feather Meal by Bacillus licheniformis S6

A two-step biotechnological process was developed using Bacillus licheniformis S6 to provide a simple and economical procedure which significantly improved feather meal nutrition value. Compared with IFM （initial feather meal） and CFM （commercial feather meal）, SFEFM （feather meal gained by solid fermentation and enzymolysis with continuous agitation） had a significant improvement （P〈0.05） in vitro digestibility, contents of oligopeptides and soluble protein released in digestive juice by pepsin- pancreatin digestion procedure, furthermore, some deficient essential amino acids in feather protein （histidine, methionine, lysine） were enhanced. Comapared with CFM, the oligopeptides released into digestive juice of ISFM （feather meal obtained by the biotechnological process described in the paper with intermittent shaking） was significantly enhanced （P〈O.05）, and its in vitro digestibility was statistically （P〉0.05） equivalent to CFM. The summary of the finding to IFM treatment and possible means of further improvements were also listed.

Effects of Different Wall-breaking Methods on Extraction of Solanesol from Stems and Leaves of Potato

In order to extract solanesol from potato stems and leaves more effectively and improve the extraction rate of solanesol, the same batch of potato stems and leaves harvested from Guyuan, Ningxia was selected as a research object, cell wall of potato stems and leaves was broken by enzymolysis with cellulase and high- speed shearing, and then reflux-extracted with 95% ethanol. Solanesol content was determined by HPLC, and extract yield was calculated. Extract yield and solanesol extraction rate was used as an index for comparison of difference between the 2 wall-breaking methods, so as to select the optimal wall-breaking method. The results showed that enzymolysis with cellulase exhibited extraction rate and extract yield of solanesol of 91.38% and 8.02%, respectively, which were better than those under high-speed shear emulsification technique. The enzymolysis wall-breaking method has the advantages of simple operation and strong feasibility.

Enzymatic pretreatment and microwave extraction of asiaticoside from Centella asiatica

The extraction of asiaticoside from Centella asiatica by enzymatic pretreatment and microwave extraction (EPME) was studied in this article. The effects of several important factors such as temperature of enzymatic pretreatment, liquid to solid ratio and microwave radiation time were investigated by quadric regression orthogonal design experiment and were analyzed by response surface. An extraction model with well forecast performance was then established. The results indicate that the optimum extraction condition was as follows: liquid to solid ratio was 36mL/g, temperature of enzymatic pretreatment was 45℃, enzymatic time was 30min, and microwave ra-diation time was

Study on Biotransformation of Traditional Chinese Herbal Medicines Complex

A newly designed enzymolysis-fermentation combined method to dramatically enhance the actives level and skin benefits of traditional Chinese herbs (TCHs) was developed, biotransformation process under optimal reaction conditions can significantly transform molecular structures and obtained fermentation extracts can deliver better skin benefits on anti-aging, hydration and whitening. Analytical results showed that the ginsenoside Rg3, total sugar, polyphenols and flavonoids in fermented extract were 2~4 times higher than unfermented extract. In-vitro tests including DPPH radical scavenging activity, tyrosinase inhibition, cells proliferation and Hyaluronan-CD44 activity, have showed significant enhancement of efficacy, inhibition rate on DPPH antioxidation as example achieved over 60% improvement, which makes traditional Chinese herbs application more feasible. It is inferred that this study potentially enables the herbs to deliver required bioefficacy in cosmetic application.

天然胡萝卜汁加工工艺的研究

以新鲜胡萝卜为主要原料,研究了生产天然胡萝卜汁的加工工艺,并通过对比试验和正交试验确定了最佳工艺参数。胡萝卜的适宜用量为20％,添加0.5％柠檬酸和0.5％异抗坏血酸,料水比1:2,在不锈钢锅中预煮10min,可保持胡萝卜汁的色泽和消除生焖味。用0.04％的果胶酶在(45±1)℃下酶解处理胡萝卜浆1.0h,可提高出汁率和胡萝卜素提取率。0.36％复合稳定剂(其中CMC:琼脂=25:11),用高压均质处理2次(均质压力为25MPa,均质温度为50℃),解决了产品沉淀问题。杀菌条件为(82±1)℃水浴中杀菌30min。

Production of Free Amino Acid and Short Peptide Fertilizer from Rapeseed Meal Fermentation Using Bacillus flexus NJNPD41 for Promoting Plant Growth

Free amino acids and short peptides(FAPS)that play an important role in plant nutrition can be extracted from rapeseed meal(RSM)by microbial enzymolysis.The enzymolytic activity of one bacterial strain isolated from RSM for the production of FAPS fertilizer(FAPSF)via solid-state fermentation,as well as its detoxification activity for isothiocyanates and oxazolidinethione,was investigated in this study.The strain NJNPD41 isolated from RSM piled-soil possessed effective proteolytic activity and was identified as Bacillus flexus on the basis of its morphological,physiological,and biochemical properties,as well as 16S rDNA analysis.Compared to the uninoculated control,inoculation with the strain NJNPD41 significantly increased the yield of total free amino acids and short peptides by 115%and decreased the content of isothiocyanates and oxazolidinethione by 53%and 60%,respectively,after solid-state fermentation.The pot and field experiments showed that FAPSF significantly promoted eggplant growth and enhanced the fruit yield and quality of eggplant compared to the control.In conclusion,this study provided a novel method for the high-value utilization of RSM to produce high-quality and low-toxicity FAPSF using the newly isolated strain B.flexus NJNPD41.

Antioxidant and ACE inhibitory activities of peptides prepared from adzuki bean by semi-solid enzymatic hydrolysis

In this study,adzuki bean peptides were prepared by semi-solid enzymatic hydrolysis(SEH)and their antioxidant and Angiotensin-I-converting enzyme(ACE)inhibitory activities and internal absorption were studied.By modifying different enzymatic hydrolysis times and enzyme additions,the optimal enzymatic hydrolysis condition for SEH could be obtained as adzuki bean powder:water=4:1(w/w),alcalase:neutrase=2:1(w/w),enzyme addition 3.75%(w/w),4 h enzymatic hydrolysis at 50℃ and pH=8.Within these hydrolysis conditions,the content of adzuki bean peptides was 11.37%,the OH−scavenging rate of 1 mg/mL peptides was 72.75%,the total antioxidant capacity was 0.88±0.02 mmol/L FeSO4,the ACE inhibition rate was 77.49%.In vivo absorption test proved that the peptides prepared by SEH were absorbed faster and exhibited higher antioxidant activity compared with liquid enzymatic hydrolysis.In conclusion,the process of SEH can improve not only the efficiency of hydrolysis,but also the antioxidant and ACE inhibitory activity as well as the internal absorption of adzuki bean peptides.

A Rapid Method for Isolating Single Cells from Apple Flesh

The identification,separation and analysis of individual living cells can be used to analyze the heterogeneity and operation mechanism of living systems.The study of fruit development is based on the extraction of active single cells.In this study,we investigated the effects of different enzymes and enzymatic hydrolysis times on the extraction of single cells from the‘Fuji’apple(Malus×domestica Borkh.‘Fuji’).The results showed that the extraction of single cells in apple flesh was a suitable method when 0.1%macerozyme was used and the enzymolysis time was 0.5 h.Fluorescent brightening agent VBL staining showed that the cell wall was intact,while fluorescein diacetate FDA and azo dye Evans blue staining indicated that the extracted single cells were active.The extracted single cells could be further used as materials for protoplast extraction.