Optimisation of freeze drying conditions for purified serine protease from mango (Mangifera indicaCv. Chokanan) peel

This study investigated the possible relationship between the encapsulation variables, namely serine protease content (9–50 mg/ml, X₁), Arabic gum (0.2–10% (w/w), X₂), maltodextrin (2–5% (w/w), X₃) and calcium chloride (1.3–5.5% (w/w), X₄) on the enzymatic properties of encapsulated serine protease. The study demonstrated that Arabic gum, maltodextrin and calcium chloride, as coating agents, protected serine protease from activity loss during freeze-drying. The overall optimum region resulted in a suitable freeze drying condition with a yield of 92% for the encapsulated serine protease, were obtained using 29.5 mg/ml serine protease content, 5.1% (w/w) Arabic gum, 3.5% (w/w) maltodextrin and 3.4% (w/w) calcium chloride. It was found that the interaction effect of Arabic gum and calcium chloride improved the serine protease activity, and Arabic gum was the most effective amongst the examined coating agents. Thus, Arabic gum should be considered as potential protection in freeze drying of serine protease.     

Purification of serine protease from mango (Mangifera Indica Cv. Chokanan) peel using an alcohol/salt aqueous two phase system

An alcohol/salt-based aqueous two-phase (ATPS) system, as a novel method of purification, was employed to purify serine proteases from mango (Mangifera Indica Cv. Chokanan) peel. The effectiveness of different parameters, such as type and concentration of alcohol (1-propanol, 2-propanol, and ethanol), type of salt (sodium citrate, potassium phosphate, and ammonium sulphate), pH, and NaCl, on the purification and selective separation of serine protease was investigated. Desirable conditions of partition coefficient (K), selectivity (S), purification factor (P), and yield (Y%) of serine protease, using ATPS, were determined. The highest partition coefficient (64.5) and selectivity (343.2) for serine protease purification value were achieved in an ATPS of 16% (w/w) 2-propanaol, 19% (w/w) potassium phosphate, and 5% (w/v) NaCl at pH 7.5. It was demonstrated that serine protease could be recovered with a yield of 96.7% and a purification factor of 11.6.     

Antioxidant and antiproliferative activities of mango (Mangifera indica L.) flesh and peel

The antioxidant and antiproliferative properties of flesh and peel of mango (Mangifera indica L.) were investigated. The cytoprotective effect of mango flesh and peel extracts on oxidative damage induced by H₂O₂ in a human hepatoma cell line, HepG2, were determined, and the underlying mechanism was examined by a single-cell electrophoresis assay (comet assay). Treatment of HepG2 cell with mango peel extract prior to oxidative stress was found to inhibit DNA damage. The free radical scavenging activities of mango flesh and peel extracts were evaluated by electron spin resonance (ESR). The mango peel extract exhibited stronger free radical scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and alkyl radicals than mango flesh extract, regardless of ripeness. Similarly, peel extract exhibited significant antiproliferative effect against all tested cancer cell lines, compared to that of flesh extract, in a dose-dependent manner. The result also showed that the antiproliferative activity of mango flesh and peel extracts correlated with their phenolic and flavonoid contents. Thus, mango peel, a major by-product obtained during the processing of mango product, exhibited good antioxidant activity and may serve as a potential source of phenolics with anticancer activity.     

Purification and characterisation of multiple forms of polygalacturonase from mango (Mangifera indica cv. Dashehari) fruit

Three multiple forms of polygalacturonase (PG) namely PGI, PGII and PGIII were isolated, purified and characterized from ripe mango (Mangifera indica cv. Dashehari) fruit. Native molecular weights of PGI, PGII and PGIII were found to be 120, 105 and 65 kDa, respectively. On SDS–PAGE analysis, PGI was found to be a homodimer of subunit size 60 kDa each while those of PGII and PGIII were found to be heterodimers of 70, 35 and 38, 27 kDa subunit size each, respectively. Three isoforms of PG differed with respect to the effect of pH, metals, reducing agents and their susceptibility towards heat. PG isoforms also differed with respect to the effect of substrate concentration on enzyme activity. PGI and PGIII exhibited inhibition at high substrate concentration while PGII did not. Km for polygalacturonic acid was found to be 0.02% for PGI.     

Low molecular weight serine protease from the viscera of sardinelle (Sardinella aurita) with collagenolytic activity: Purification and characterisation

A new low molecular weight (LMW) serine-protease from sardinelle (Sardinella aurita) viscera was purified using ammonium sulphate precipitation and Sephadex G-100 gel filtration, with a 3.82-fold increase in specific activity. The molecular weight of the enzyme was estimated to be 14.2 kDa by SDS-PAGE. The optimum pH and temperature for the enzyme activity were around pH 8.0 and 60 °C, respectively. The purified protease was strongly inhibited by phenylmethylsulphonyl fluoride, a serine-protease inhibitor, and soybean trypsin inhibitor. The N-terminal amino acid sequence of the first 10 amino acids of the purified protease was APVQPCVVVI. This sequence showed low homology with several peptidases, suggesting that the enzyme is a new protease. Interestingly, the protease was found to cleave collagen type I and hydrolyze succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (sAAPFpna), an amide substrate of chymotrypsin. Our findings indicate that the S. aurita protease is a new LMW enzyme with collagenolytic activity.     

Religiosin B, a milk-clotting serine protease from Ficus religiosa

A novel milk-clotting serine protease, named religiosin B, is purified from Ficus religiosa. The molecular mass of the protein is 63,000 with pI value of pH 7.6. The proteolytic activity of the enzyme is strongly inhibited by phenylmethanesulfonyl fluoride (PMSF) and chymostatin. Religiosin B acts optimally at pH 8.0-8.5 and temperature 55 °C. The molar absorption coefficient of the enzyme is 149,725 M⁻¹cm⁻¹ with 23 tryptophan, 15 tyrosine and 7cysteine residues per molecule of the enzyme. The enzyme shows broad substrate specificity with natural as well as synthetic substrates. Religiosin B is highly stable against denaturants and metal ions as well as over a wide range of pH and temperature. The de novo sequencing confirms the novelty of the enzyme. In addition to its high milk-clotting ability, it could be used in the cheese industry, as well as other food and biotechnological industries.     

Physico-chemical and antioxidant properties of four mango (Mangifera indica L.) cultivars in China

Four principal mango cultivars (Tainong No.1, Irwin, JinHwang and Keitt) grown in southern China were selected, and their physico-chemical and antioxidant properties were characterized and compared. Of all the four cultivars, Tainong No.1 had highest content of total phenols, ρ-coumaric acid, sinapic acid, quercetin, titratable acidity, citric acid, malic acid, fructose, higher antioxidant activities (DPPH, FRAP) and L^∗, lower pH, PPO activity and individual weight. Keitt mangoes showed significantly (p < 0.05) higher contents of β-carotene, ρ-hydroxybenzoic acid, sucrose, total sugar, total soluble solid, catechin, succinic acid and higher PPO activity. JinHwang mangoes exhibited significantly (p < 0.05) higher individual weight and PPO activity, but had lower content of total phenols, β-carotene and lower antioxidant activity. Principal component analysis (PCA) allowed the four mango cultivars to be differentiated clearly based on all these physico-chemical and antioxidant properties determined in the study.     

菠萝蜜果皮黄酮提取工艺优化及比较

为筛选适合菠萝蜜果皮黄酮的提取方法,以得率为评价指标,应用酶法和有机溶剂浸提法提取其果皮中的黄酮类化合物,在单因素基础上,通过正交实验,优化提取工艺,并对两种提取方法结果进行比较评价。结果表明,酶法提取菠萝蜜果皮黄酮最佳工艺条件为:果胶酶用量300 U/g、乙醇浓度80%(v/v)、温度55℃、p H5.5、底物质量浓度45 g/L,时间2.0 h;有机溶剂浸提最佳工艺条件为:乙醇浓度80%(v/v),温度55℃,料液比1∶20(g/m L),时间2.5 h;酶法提取黄酮类化合物得率和纯度分别为4.98%、12.60%,高于有机溶剂浸提法的3.05%、8.92%,提取物的抗氧化性强于有机溶剂浸提法,说明酶法提取菠萝蜜果皮黄酮优于传统有机溶剂浸提法。      

石榴皮原花青素的微波提取工艺及其稳定性研究

采用微波法提取野生石榴皮中的原花青素,同时考察光照、温度、金属离子、PH及食品添加剂对原花青素稳定性的影响。通过单因素试验和正交试验得出微波提取石榴皮原花青素的最佳工艺条件为：乙醇浓度80％、料液比1：50、提取温度65℃、微波功率500W、提取时间120s、提取2次,原花青素的得率为2．62％。稳定性研究表明,光照条件下色素稳定性较差;高温对色素有降解作用,60℃以下稳定性良好;pH值为5时色素稳定性好;金属离子Cu^2＋和Al^3＋对原花青素的稳定性影响显著;苯甲酸对原花青素稳定性影响显著。     

Optimisation of extraction conditions for recovering carotenoids and antioxidant capacity from Gac peel using response surface methodology

The peel of Gac fruit is regarded as waste product in the processing of Gac although it contains high level of carotenoids and possesses a significant antioxidant capacity. This study optimised the extraction yields of carotenoids and antioxidant capacity from Gac peel. Different organic solvents were examined to determine the most suitable solvent for the extraction. The extraction conditions including time, temperature and solvent–solid ratio were then optimised for maximising extraction yields of carotenoids and antioxidant capacity from Gac peel using response surface methodology. Ethyl acetate was identified as the most suitable solvent. The optimal extraction time, temperature and solvent–solid ratio were 150 min, 40.7 °C and 80 mL g^?1, respectively. The carotenoid extraction yield and the antioxidant capacity extraction yield were 271 mg/100 g DW and 737 μm TE/100 g DW, respectively. Thus, the extraction using ethyl acetate with the ratio of 80:1 (mL solvent per g Gac peel) for 150 min at 40.7 °C is suggested for recovering carotenoids and antioxidant capacity from Gac peel.     

Ultrasound-assisted extraction of polyphenols (flavanone glycosides) from orange (Citrus sinensis L.) peel

The present study reports on the extraction of polyphenols especially flavanones from orange (Citrus sinensis L.) peel by using ethanol as a food grade solvent. After a preliminary study showing that the best yield of extraction was reached for a particle size of 2 cm², a response surface methodology (RSM) was launched to investigate the influence of process variables on the ultrasound-assisted extraction (UAE) followed by a central composite design (CCD) approach. The statistical analysis revealed that the optimised conditions were a temperature of 40 °C, a sonication power of 150 W and a 4:1 (v/v) ethanol:water ratio. The high total phenolic content (275.8 mg of gallic acid equivalent/100 g FW), flavanone concentrations (70.3 mg of naringin and 205.2 mg of hesperidin/100 g FW) and extraction yield (10.9 %) obtained from optimised UAE proved its efficiency when compared with the conventional method. Furthermore, the antioxidant activity determined by the DPPH and ORAC tests confirmed the suitability of UAE for the preparation of antioxidant-rich plant extracts.     

从柑桔皮中超声提取橙皮苷

从柑桔皮中连续提取色素、果胶和橙皮苷。浸提果胶后的滤渣,在超声波作用下,利用饱和Ca(OH)2碱液浸提盐酸酸析得到橙皮苷。最佳工艺条件为:碱提pH12.5,液料比10:1,超声时间60min,酸析pH4.7,浸提级数3级。另外,考察了重结晶法精制对橙皮苷纯度的影响,平均纯度为94.96%,提高14.72%。红外光谱、高效液相色谱等方法表征实验产品是橙皮苷。     

超声波辅助提取菠萝皮多酚物质的研究

研究了超声波辅助提取菠萝多酚物质的工艺,对影响多酚浸提率的因素进行了单因素试验和正交试验,确定了最佳工艺条件为:乙醇浓度60%,浸提温度45℃,功率200W,浸提时间60min,液料比15.0∶1.0。在此条件下,浸提率可达18.48g/kg。     

微波辅助提取石榴皮多酚工艺研究

研究从石榴皮中微波辅助提取多酚的工艺.采用单因素试验及正交试验,考察不同条件下石榴皮多酚提取率.结果表明,提取溶剂乙醇体积分数、微波功率、提取时间得料液比均对石榴皮多酚提取率有显著影响;石榴皮多酚的最佳提取条件为50％(体积分数)乙醇作溶剂,微波功率300W,提取时间120s,料液比1∶35(m∶V).该条件下提取石榴皮多酚,提取率可达21.41％.     

Multiple forms of polygalacturonase from mango (Mangifera indica L. cv Alphonso) fruit

Polygalacturonase (PG) from mango pulp revealed three isoforms (I, II, III) upon ion exchange and gel filtration chromatography, each having an abundance of 68%, 6% and 26%, and molecular weights (M_r) 40, 51 and 45 kDa, respectively. The pH optimum for the isoforms was between 3 and 4. PG-I was stable over a wide pH range (4–7.5) unlike PG II and III, which were stable at pH 4 and 5, respectively. The optimum temperature was around 40 °C for all the three isoforms. Their apparent K_m for pectic acid was in the range 0.22–0.25 mg ml⁻¹. The V_max for PG I, II and III was 5.7, 3.6 and 4.4 μmol GalA equivalent h⁻¹, respectively. Cd²⁺, Cu²⁺ and Fe²⁺ and EDTA inhibited whereas GalA, Gal, Fuc, Rha and Ara stimulated PG-I activity, in particular. The major endogenous substrates for mango PG were identified to be two rhamnogalacturonans varying in their sugar ratio. These results are discussed in the light of pectin dissolution in vivo in ripening mango.     

微波辅助提取苹果皮中的多酚类物质

以苹果皮为原料提取苹果多酚,对不同提取剂的提取效果进行了比较;通过单因子试验和正交试验对微波辅助提取多酚的工艺条件进行了探讨,研究了溶剂浓度、微波功率、微波处理时间以及料液比对多酚提取的影响;并对微波辅助提取和常规法提取的效果进行了比较。结果表明：综合考虑提取效果及食品安全性,选择乙醇作为提取剂为宜;微波辅助提取的最佳工艺条件为乙醇浓度70%、微波功率210W、微波时间25s、料液比（w/v）1∶5;微波辅助提取法提取物中原花色素含量0.3668mg/ml,常规法提取物中原花色素含量0.2552mg/ml,微波辅助提取效果优于常规法提取。     

Quantification of gallic acid and ellagic acid from longan (Dimocarpus longan Lour.) seed and mango (Mangifera indica L.) kernel and their effects on antioxidant activity

Gallic acid (GA) and ellagic acid (EA) have been identified in longan seed and mango kernel by the use of reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with photodiode array detection (DAD). The ethanolic extract of longan seed contained 23.3 and 156 mg/100 seeds of GA and EA, respectively. The ethanolic extracts of mango kernel contained approximately 87% more GA than the longan seed ethanolic extracts but about 32% less EA. After heat treatment and acid hydrolysis, mango kernel had higher concentrations of GA and EA, contributing to more potent antioxidant activity. The results demonstrated rich sources of GA and EA in longan seed and mango kernel which might provide a novel source of these natural antioxidants.     

秦巴山区柑橘皮多糖的提取工艺及稳定性研究

以秦巴山区柑橘皮为原料,采用微波预处理-热水浸提法,在不同料液比、微波时间、浸提时间和浸提温度下探讨柑橘皮多糖的提取工艺,并对其多糖在不同pH和常见的一些氧化剂、还原剂、食品添加剂、金属离子等条件下的稳定性进行研究。实验结果表明,柑橘皮多糖的最佳提取条件:料液比1∶40（g/mL）、微波时间90s、浸提时间1.5h、浸提温度80℃,多糖提取率为39.25%;柑橘皮多糖在酸性、还原剂介质中稳定性较好,在氧化剂、碱性、苯甲酸钠、柠檬酸和K+、Na+、Ca2+、Al3+等金属离子溶液中的稳定性相对较差。      

Rheological and high gelling properties of mango (Mangifera indica) and ambarella (Spondias cytherea) peel pectins

Ambarella and mango peels are good sources of pectins (15–20%), with high degree of methylation (60–78%) and high molar masses. Ambarella and mango ( Améliorée and Mango varieties) peel pectins were extracted using HCl or oxalic acid/ammonium oxalate (OAAO). Purified pectins were analysed for their flow behaviour and phase diagrams were established at pH 3 as sucrose vs. pectin concentration. The gelation kinetics and mechanical spectra of these pectin gels were studied and compared to those of commercial citrus (lime) pectins. At a concentration of 1% (w/v), all pectic solutions had a shear thinning behaviour but at 0.6% (w/v), only OAAO-extracted pectins exhibited such behaviour. Phase diagrams showed that at pH 3, gelation of OAAO mango extracted pectins was possible at low polymer concentration (0.2%; w/w) for a sucrose concentration of 60% (w/w). OAAO-extracted pectins exhibited a higher gelling ability than HCl-extracted ones. Sucrose (45–50%) and pectin (0.2–0.6%) concentration had a deep impact on the gel strength. Our results enable to conclude that the OAAO extraction from mango and ambarella peels allowed the recovery of pectins that exhibit high gelling properties.     

响应面法优化超声波辅助提取猕猴桃果皮多酚工艺研究

采用超声波辅助提取猕猴桃果皮多酚,并利用响应面法对多酚提取工艺进行优化。在单因素实验的基础上,采用四因素三水平的响应面实验优化设计,研究超声波功率、提取时间、提取温度、液料比对多酚提取量的影响。结果显示最佳提取工艺条件为:超声波功率384.00 W,提取时间30 min,提取温度65.00℃,液料比23.00 m L/g,多酚提取量的实验值为（28.10±0.38）mg GAE/g,与理论预测值（28.14 mg GAE/g）相差不大。通过体外1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除力测定多酚的抗氧化性,并且得到猕猴桃果皮多酚的EC50值为0.13 mg/m L,说明提取的多酚具有很好的抗氧化性。