乳酸菌发酵法富集辣木叶中γ-氨基丁酸的研究

【目的】利用乳酸菌为发酵菌株,对辣木叶中的γ-氨基丁酸(γ-aminobutyric acid,GABA)进行发酵富集。【方法】对2株报道产GABA的商业菌株及两株实验室筛选产GABA的菌株进行筛选,得到最适发酵菌株。对辣木粉添加量、谷氨酸钠(MSG)添加量、初始pH、发酵温度、发酵时间进行单因素实验,并选择MSG添加量、初始pH、发酵温度,发酵时间进行正交实验,得出最佳发酵条件。【结果】4种乳酸菌产GABA能力大小依次为植物乳杆菌LK-1植物乳杆菌S35耐久肠球菌干酪乳杆菌LK-1,利用植物乳杆菌LK-1发酵辣木叶,GABA的含量可高达152 mg/L。经单因素及正交实验,确定GABA的最佳发酵条件为辣木叶粉添加量为质量分数4%,MSG添加量为4g/L,初始pH6.5,发酵温度为35℃,发酵时间72h。在此条件下发酵辣木液GABA质量浓度可高达209mg/L,比未发酵的辣木液提高1.45倍。【结论】利用植物乳杆菌LK-1发酵辣木叶能大大提高GABA的含量。     

木瓜蛋白酶在制取青鳞鱼可溶性蛋白中的应用

研究了木瓜蛋白酶的酶解温度、酶浓度、酶解时间、加水量、pH值对青鳞鱼可溶性蛋白得率的影响 ,探讨了α 氨态氮含量与青鳞鱼可溶性蛋白得率的关系。结果表明 :温度小于 5 0℃ ,随着酶解温度的提高、酶浓度的增加、酶解时间的延长青鳞鱼可溶性蛋白得率及α 氨态氮的含量均增加 ,α 氨态氮含量增加比可溶性蛋白得率的增加明显 ;加水量的增加 ,可溶性蛋白得率增加 ;pH值对水解效果及可溶性蛋白得率影响不大。综合考虑 ,最佳水解条件为 :温度 5 0℃ ,酶浓度为 1 5 0 0u/g ,酶解时间 80min ,m原料 :m水 =1∶2 ,pH 6 8。酶解所得可溶性蛋白经氨基酸分析仪检测 ,其氨基酸总量为 49 47mg/mL ,必需氨基酸占氨基酸总量的 41 0 % ,游离氨基酸含量为 6 3mg/mL ,肽含量为 87 2 %。     

利用小杂鱼,低值鱼提取浓缩鱼蛋白

以小杂鱼、低值鱼为原料,采用溶剂萃取法研究了溶剂种类温度、时间和溶剂量对鱼蛋白提取过程中脱水、脱脂、脱色和脱腥的影响,筛选出提取浓缩鱼蛋白最佳工艺条件。结果表明：在该工艺条件下制得的浓缩鱼蛋白提取其蛋白质含量达９２．２８％,脂肪含量仅为０．０９２％。     

利用小杂鱼、低值鱼提取浓缩鱼蛋白

以小杂鱼、低值鱼为原料,采用溶剂萃取法研究了溶剂种类、温度、时间和溶剂量对鱼蛋白提取过程中脱水、脱脂、脱色和脱腥的影响,筛选出提取浓缩鱼蛋白的最佳工艺条件。结果表明,在该工艺条件下制得的浓缩鱼蛋白,其蛋白质含量高达９２．２８％,脂肪含量仅为０．０９２％。     

波吉卵囊藻蛋白质提取方法比较及提取条件优化

【目的】探究基于考马斯亮蓝染色法的波吉卵囊藻(Oocystis borgei)蛋白质提取方法,以快速测定波吉卵囊藻的蛋白质含量。【方法】以凯氏定氮法为基准,以考马斯亮蓝法测定经反复冻融法、超声波破碎法和碱提法提取的波吉卵囊藻(鲜品)蛋白含量,比较各提取方法的提取效果;通过单因素和响应面实验,考察碱提法中提取温度、NaOH浓度和提取时间对波吉卵囊藻蛋白提取率的影响。【结果】碱提法显著优于其他提取方法(P 0.01);碱提法中,各因素对波吉卵囊藻蛋白提取率的影响显著性由大到小依次为提取温度、NaOH浓度、提取时间;最佳提取条件:提取温度74.5℃、NaOH浓度0.35mol/L、提取时间69min,此时蛋白得率为1.726%,与模型预测值1.731%接近,以凯氏定氮法测定的藻体蛋白质含量为基准,蛋白提取率高达99.20%。【结论】碱提法是波吉卵囊藻(鲜品)蛋白的最佳提取方法,采用响应面法建立的模型可较好地预测提取温度、NaOH浓度和提取时间对波吉卵囊藻蛋白提取率的影响。     

木瓜蛋白酶在制取青鳞鱼可溶性蛋白中的应用

研究了木瓜蛋白酶的酶解温度，酶浓度，酶解时间，加水量，pH值对青鳞鱼可溶性蛋白得率的影响，探讨了α－氨态氮含量与青鳞鱼与可溶性蛋白得率的关系。结果表明：温度小于50℃，随着酶解温度的提高，酶浓度的增加，酶解时间的延长青鳞鱼可溶性蛋白得率及α－氨态氮的含量均增加，α＝氨态氮含量增加比可溶性蛋白得率的增加明显；加水量的增加，可溶性蛋白得率增加；pH值对水解效果及可溶性蛋白得率影响不大。综合考虑，最佳水解条件为：温度50℃，酶浓度为1500u/g，酶解时间80min,m原料：m水＝1：2，pH6.8。酶解所得可溶性蛋白经氨基酸分析仪检测,其氨基酸总量为49.47mg/mL，必需氨基酸占氨基酸总量的41．0％，游离氨基酸含量为6.3mg/mL，肽含量为87．2％。     

响应面法优化波吉卵囊藻多糖提取条件

【目的】探究波吉卵囊藻(Oocystis borgei)多糖提取的最优条件。【方法】通过单因素和响应面实验,考察不同提取温度、氢氧化钠浓度、提取时间对波吉卵囊藻多糖提取率的影响。【结果】各因素对波吉卵囊藻多糖提取率的影响显著性表现为氢氧化钠浓度提取温度提取时间;最佳提取条件:提取温度58.5℃、氢氧化钠浓度0.305mol/L、提取时间68 min,此时提多糖取率为11.86%,与模型预测值11.93%接近。【结论】采用响应面法建立的模型可以较好地预测提取温度、氢氧化钠浓度和提取时间对波吉卵囊藻多糖的提取率的影响。     

酶解法提取鱼油的工艺参数优化

利用蛋白酶酶解法从黄鳍金枪鱼(Thunnus albacares)加工的下脚料———鱼头中提取鱼油。以鱼油提取率和感官特征为指标,通过正交优化实验设计,获得了胰蛋白酶提取鱼油的最佳酶解工艺参数:酶解温度45℃,酶添加量1.5%,料液质量比1∶1,酶解时间4 h,酶解pH 8。在该条件下,鱼油的提取率为4.34%,理化指标除过氧化值外均达到SC/T3502-2000的粗鱼油二级标准,多不饱和脂肪酸总含量高达38.47%,其中DHA和EPA的含量分别为23.63%和4.84%。     

超声波辅助提取河豚毒素的研究

以月腹刺鲀(Gastrophysus lunaris)卵巢为原料,利用超声波辅助提取河豚毒素(Tetrodotoxin,TTX)。通过单因素实验及二次回归旋转组合设计实验研究液料比、超声温度、超声时间等因素对提取率的影响,确定超声波辅助提取河豚毒素的最佳工艺条件。结果表明:在实验范围内影响河豚毒素得率的各因素大小依次为液料体积质量比、超声温度、超声时间;超声波辅助提取河豚毒素的最佳工艺条件为液料体积(mL)质量(g)比3.03∶1,超声温度39℃,超声时间22 min,该工艺条件下提取河豚毒素得率为63.57μg/g。     

酶解法提取鱼油的工艺参数优化

利用蛋白酶酶解法从黄鳍金枪鱼（Thunnus albacares）加工的下脚料——鱼头中提取鱼油。以鱼油提取率和感官特征为指标，通过正交优化实验设计，获得了胰蛋白酶提取鱼油的最佳酶解工艺参数：酶解温度45℃，酶添加量1．5％．料液质量比1：1，酶解时间4h，酶解pH8。在该条件下．鱼油的提取率为4．34％，理化指标除过氧化值外均达到SC／T3502-2000的粗鱼油二级标准。多不饱和脂肪酸总含量高达38．47％,其中DHA和EPA的含量分别为23．63％和4．84％。     

Ultrasonic-assisted condensation of chitosan with salicylaldehyde

In order to synthesize an improved adsorbent for heavy metal ions, we studied the condensation reaction of chitosan with salicylaldehyde in ethanol to form a Schiff base. The effect irradiating the reaction using an ultrasonic liquid processor was contrasted with conventional methods. The IR spectra of condensed chitosan prepared by the two methods showed that their molecular structures were identical. The reaction conditions, including solvents, ultrasonic power density and irradiation time, pH, and reactant ratio, were optimized by orthogonal design. A shorter reaction time and a higher product yield were obtained using ultrasonic-assisted synthesis compared with the traditional method. A condensation degree of 89.63% was achieved using the optimized conditions: i.e. ultrasonic irradiation at 180 W for 60 min; 95% ethanol as the solvent, pH 4.0, and salicylaldehyde:chitosan ratio of 6:1.     

Preparation by enzymolysis and bioactivity of iron complex of fish protein hydrolysate （Fe-FPH）from low value fish

Preparation of Fe^2＋ chelate of fish protein hydrolysate （Fe-FPH） obtained from low value fish proteins was introduced and its bioactivity was studied by compound enzymolysis. The optimum conditions for hydrolysate chelating Fe^2＋ are DH （degree of hydrolysis） at 5%, pH 7.0, 20℃ and 15 min chelating time for FM （material not being defatted）. Four types of Fe-FPH including CA （deposit after chelating）, CB （deposit in 50% of absolute ethanol solution）, CC （suspended deposit in 80% of absolute ethanol solution）, and CD （bottom deposit in 80% of absolute ethanol solution） were fractionated with absolute ethanol from FM. Structural analysis through infra-red spectrum revealed that Fe^2＋ was combined strongly with amino-group and carboxyl-group in each chelate and each Fe^2＋ could form two five-member ring structures. All of the four chelates were shown more significant antioxidative activity and can be used as natural hydrophobic and hydrophilic antioxidant. Among all the chelates, the CB possesses the most effective antioxidative activity at 92% as high as that of a-tocopherol. Among all Fe-FPHs, only CD showed the most effective antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Bacillus subtilis and can be used as natural antibacterial. It provides a more effective way for utilization of low value fish proteins and key information of Fe-FPH as additive in food industry.     

Role of organic acids in desorption of mercury from contaminated soils in eastern Shandong Province, China

Batch experiments were conducted to study the effects of low-molecular-weight organic acids(LMWOAs) on desorption of Hg from two kinds of soils in the eastern Shandong Province,China.Of all LMWOAs,oxalic and citric acids were chosen as the representatives in this experiment because they are most common in soils.Desorption of Hg increases with the increase of extracting time,until the highest desorption amount at 6-8 h,and then declines.It indicates that timeliness is very important when organic acids are used for phytoremediation in contaminated soils.When the concentrations of organic acids increase from 1.0 mmol/L to 10.0 mmol/L,Hg desorption increases significantly regardless of oxalic or citric acid.Compared with oxalic acid,citric acid is more effective to enhance the desorption of Hg,especially from fluvo-aquic soil.With the increase of initial pH value from 3 to 8,Hg desorption decreases directly in the presence of oxalic acid.However,Hg desorption first decreases,then increases,and eventually decreases again in the presence of citric acid due to the variation of competitive ability between chelating of citric acid and adsorption of soil in different pH values.Citric acid showed greater ability to release Hg from soils than oxalic acid.According to the results of the present study,it is found that the bioavailabilities of heavy metal can be improved with selecting suitable types and concentrations of organic acid amendment and reasonable soil condition.     

不同方式提取的近江牡蛎糖蛋白的体外抗氧化活性

比较研究了水浸提、盐液浸提及超声辅助盐液浸提的近江牡蛎糖蛋白粗提物对羟自由基、DPPH自由基、超氧阴离子自由基的清除效果。结果表明:水浸提、盐液浸提及超声辅助盐液浸提的糖蛋白中可溶性蛋白与总糖含量之比分别为1.83∶1、4.49∶1和2.51∶1;3种糖蛋白粗提物均具有一定的清除羟自由基、DPPH自由基和超氧阴离子自由基的能力;它们对羟自由基的清除能力高于对DPPH自由基和超氧阴离子自由基;盐液浸提物清除超氧阴离子的能力最弱;超声辅助盐液浸提的糖蛋白粗提物清除羟自由基、超氧阴离子自由基及DPPH自由基的能力比水浸提和盐液浸提的糖蛋白的能力强。     

Comparative study on the protease inhibitors from fish eggs

The protease inhibitor was purified from five different fish eggs. The molecular weights of Pacific herring, chum salmon, pond smelt, glassfish, and Alaska pollock egg protease inhibitors were 120, 89, 84.5, 17, and 16.8kDa, respectively. The specific inhibitory activity of glassfish egg protease inhibitor was the highest followed by those of Pacific herring and Alaska pollock in order. The specific inhibitory activity and purity of glassfish egg protease inhibitor were 19.70 U mg^- 1 protein and 164.70 folds of purification, respectively. Glassfish egg protease inhibitor was reasonably stable at 50 - 65℃ and pH 8, which was more stable at high temperature and pH than protease inhibitors from the other fish species. Glassfish egg protease inhibitor was noncompetitive with inhibitor constant （Ki） of 4.44 nmol L^-1     

总状蕨藻多糖提取工艺的研究

采用酶法提取与热水抽提相结合的方法研究了总状蕨藻多糖的提取工艺,结果显示,采用固液质量比1∶10,蛋白酶用量136.5 U/g,于60℃下酶法提取2.5 h后,再将固液质量比调至1∶20,在温度95℃条件下继续进行热水提取2.5 h,多糖得率为6.7%,比单用热水提取法提高了131.0%。     

螺旋藻制取叶绿素铜钠盐

用乙醇提取螺旋藻中的叶绿素,再用５％ＮａＯＨ溶液皂化、石油醚萃取,在酸性环境中用１０％ＣｕＳＯ４溶液实现铜镁置换,沉淀物经丙配溶解后,加２％ＮａＯＨ的乙醇溶液成盐,过滤,水洗,真空干燥,制成叶绿素铜钠盐．制品的质量达到ＧＢ３２６２－８２的要求．提取率为６４．７％,制品得率占藻粉１．１％。着重对铜镁置换的条件、制品耐酸、耐热、耐光等性能进行了对比试验,结果表明整个工艺是切实可行的．     

Optimization of preparative separation and purification of total polyphenols from Sargassum tenerrimum by column chromatography

Polyphenols from the ethanol extracts of Sargassum tenerrimum (ST) with potent antiallergic effects were studied to optimize separation process through column chromatography. The adsorption and desorption characteristics of three widely used adsorbents: macroporous resin, silica gel, and polyvinylpolypyrrolidone (PVPP), were critically evaluated respectively and studied for the optimization of preparative separation of polyphenols. Static operations on these adsorbents showed that macroporous resin had the best adsorption and desorption capability among the three adsorbents. Dynamic adsorption and desorption with macroporous resin packed column were also conducted to optimize the parameters such as: with the optimal values shown in brackets, the concentration of extract solution (4 times diluted), pH value (6–7), adsorption speed (3 BV h⁻¹, bed volumes/per hour), concentration of ethanol (80%), elution speed (3 BV h⁻¹) and elution volume (7 BV). The chromatographic process so optimized gave a purity of 62.43% from the crude polyphenols, providing a promising basis for large scale preparation of bioactive polyphenols upon further scaling up tests.     

Optimum production and characterization of an acid protease from marine yeast <Emphasis Type="Italic">Metschnikowia reukaufii</Emphasis> W6b

The marine yeast strain W6b isolated from sediment of the South China Sea was found to produce a cell-bound acid protease. The crude acid protease produced by this marine yeast showed the highest activity at pH 3.5 and 40 °C. The optimal pH and temperature for the crude acid protease were in agreement with those for acid protease produced by the terrestrial yeasts. The optimal medium of the acid protease production was seawater containing 1.0% glucose, 1.5% casein, and 0.5% yeast extract, and the optimal cultivation conditions of the acid protease production were pH 4.0, a temperature of 25 °C and a shaking speed of 140 rmin⁻¹. Under the optimal conditions, 72.5 UmL⁻¹ of acid protease activity could be obtained in cell suspension within 48 h of fermentation at shake flask level. The acid protease production was induced by high-molecular-weight nitrogen sources and repressed by low-molecular-weight nitrogen sources. Skimmed-milk-clotting test showed that the crude acid protease from the cell suspension of the yeast W6b had high skimmed milk coagulability. The acid protease produced by M. reukaufii W6b may have highly potential applications in cheese, food and fermentation industries.