嗜碱芽孢杆菌产环糊精葡萄糖基转移酶发酵条件的优化

对一株嗜碱芽孢杆菌产环糊精糖基转移酶的发酵条件进行了研究。利用单因素试验和正交试验获得该菌株产环糊精糖基转移酶的最佳条件为：接种量3％；培养温度30℃；pH10．5；发酵培养基的组成为玉米粉2％，酵母膏1．5％，玉米浆5％；250ml三角瓶装液量为30ml；270r/min振荡培养3d，其发酵液酶活可达5400U／ml左右。10L罐发酵时酶活可达5820U／ml。     

克雷伯杆菌甘油脱氢酶的分离纯化及性质

在有氧条件下,利用Q Sepharose Fast Flow离子交换层析和Blue Sepharose CL 6B亲和层析提纯克雷伯杆菌胞内甘油脱氢酶.酶的纯化倍数和回收率分别为 32.61 倍和 5.83%.通过SDS PAGE电泳测得该酶亚基的相对分子质量约为 34 000.该酶最适表观反应温度和最适反应pH值分别为60 ℃和11.在30 ℃以下及pH值10~12时,该酶具有良好的稳定性.在45 ℃和pH值11条件下,该酶以甘油和NAD 为底物的米氏常数Km分别为 0.75 mmol/L和 0.12 mmol/L.甘油脱氢酶对甘油的生理反应活性最大,对其它醇类如 1,2 丙二醇、乙二醇也有氧化能力.NH4 和Na 对酶有显著激活作用.巯基保护剂可明显地提高酶的活力.     

克雷伯杆菌利用生物柴油副产物甘油生产氢气和1,3-丙二醇的研究

以Klebsiella pneumoniaeDSM2026为出发菌株,通过紫外线诱变,选育得到能耐较高浓度生物柴油副产物甘油生产H2和1,3-丙二醇(1,3-PD)的菌株21株,命名为Kp1~Kp21。通过比较,Kp8菌株产量最高,1,3-PD和H2产量分别达到0.36 g/50 mL和0.99 mmol/50 mL,比出发菌株分别提高了3.5倍和4.2倍。对Kp8菌株发酵条件进行优化,得到最佳培养条件为pH 7.0,培养温度37℃,接种量10%(v/v),废甘油浓度为30 g/L。在该条件下H2产量为1.0 mmoL/50 mL,1,3-PD产量为7.5 g/L,甘油转化率为83.3%。     

用克雷伯氏菌批式流加发酵法生产1,3-丙二醇

通过对克雷伯氏菌在 7L发酵罐中厌氧间歇发酵甘油生产 1,3 丙二醇的实验研究 ,建立了一种与 pH调节相偶联的批式流加甘油发酵策略。考察了不同甘油维持浓度条件下的流加方式及不同培养方式对 1,3 丙二醇产率的影响。结果表明 ,甘油质量分数维持在 2 %的流加方式有利于 1,3 丙二醇的发酵生产 ,其在 30 5h内消耗甘油 2 80 g ,得到 1,3 丙二醇152 6 g ,摩尔转化率 6 5 5% ,生产强度 0 91g/L·h     

木糖发酵高产2,3-丁二醇菌株的选育

以肺炎克雷伯氏菌(Klebsiella pneumoniae)CICC10011为出发菌株,进行紫外诱变选育。通过2,3-丁二醇抗性和产酸能力对诱变菌株进行初筛,发酵后23,-丁二醇产量检测进行复筛,获得6株高产2,3-丁二醇的肺炎克雷伯氏菌,其中产量最高的一株诱变菌U3-17,与出发菌株相比,产量提高了21.5%。在摇瓶批式补料发酵中,诱变株U3-17的23,-丁二醇产量达49.3 g/L。     

利用来源于Paenibacillus macerans的α-CGTase突变体Y89D生产α-环糊精

对α- 环糊精葡萄糖基转移酶(α-CGTase)突变体Y89D 制备α- 环糊精的影响因素进行初步研究。其因素包括淀粉种类(马铃薯淀粉、玉米淀粉、木薯淀粉、可溶性淀粉)、加酶量、反应时间、pH 值、有机溶剂(乙醇、异丙醇、正丁醇、正癸醇)和温度。结果表明：选用5g/100mL 马铃薯淀粉、pH5.0、温度30℃、加酶量控制在 每克淀粉5U 左右,反应体系中加入体积分数5% 的正癸醇,反应6h 后,淀粉总转化率可达70%,其中α- 环糊精在产物中质量分数约为85%,转化产物中含有少量β- 环糊精(15%),而极少生成γ- 环糊精。因此,α-CGTase突变体Y89D 在制备α- 环糊精工艺中具有很好的工业化应用前景。     

Inhibitory effects of various essential oils and individual components against extended-spectrum beta-lactamase (ESBL) produced by Klebsiella pneumoniae and their chemical compositions

In the current study, in vitro inhibitory activity of several essential oils obtained from the cultivated plants, Foeniculum vulgare, Mentha piperita and M. spicata, Ocimum basilicum, Origanum majorana, O. onites, O. vulgare, Satureja cuneifolia, and a number of individual essential oil components of terpene and aromatic types were screened against 10 isolated strains of Klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) enzyme, which makes this microorganism quite resistant against the antibiotics: trimetoprime-sulfametoksazol, sulbactam-ampicilin, clavulonate-amoxicilin, ceftriaxon, cefepime, imipenem, ceftazidime, tobramicine, gentamisine, ofloxacin, and ciprofloksasin. All of the essential oils and the components exerted a remarkable inhibition ranging between 32 and 64 μg/mL against all of these strains as strong as the references (ampicilin and oflaxocin) inhibiting at 32 μg/mL. Besides, chemical compositions of the essential oils were elucidated by gas chromatography-mass spectrometry (GC-MS). The essential oils and the pure components widely found in essential oils screened herein have shown remarkable inhibition against ESBL-producing K. pneumoniae strains, which leads to the suggestion that they may be used as food preservatives for this purpose. Practical Application: The essential oils obtained from Foeniculum vulgare, Mentha piperita and M. spicata, O.cimum basilicum, Origanum majorana, O. onites, O. vulgare, and Satureja cuneifolia as well as common essential oil components have shown notable inhibitory effects against 10 isolated strains of Klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) enzyme and they might be used as food preservative or ingredient.     

R‐Index Measure of Microencapsulated Tributyrin in Gamma‐Cyclodextrin Influenced by Drying Method

Microencapsulation is commonly used in the food industry for a variety of purposes including added ingredient functionally and taste‐masking for those ingredients with negative sensory qualities. Tributyrin (TB), a source intestinally‐essential butyric acid, possesses negative aroma (cheesy, fecal) and taste (bitter) qualities. This has significantly limited its use in food applications for the potential improvement of intestinal health. Utilizing spray drying and low‐temperature oven drying, microcapsules containing TB were produced using whey (WPI), WPI and inulin, and gamma‐cyclodextrin (GCD). To determine how microcapsule formulation and drying method affected the perception of TB relative to a control, microencapsulated and free TB were added to an infant formula system and evaluated using the rating method to determine R‐index measures. Pooled R‐index measures (α = 0.01, 2‐tailed, and n = 170) indicated that the only microcapsule not significantly different from the control (R‐index below 57.95%) was the GCD and TB oven dried (GCT OD) microcapsule. All other WPI, WPI–inulin, and GCD and TB spray‐dried (GCT SD) microcapsules were all significantly different from the control. Average individual R‐index results indicated that all microcapsules in infant formula, except for GCT OD, were significantly different (P < 0.01) from the control formula but not from free TB. Spray drying may create microcapsules with surface TB and disturb the GCD–TB complex, allowing free, and surface TB to be perceived by the panelists. The GCT OD microcapsule has the potential to be used for the potential oral treatment of intestinal disorders in functional food applications without the negative sensory qualities of TB.     

Synthesis and Characterization of Nano‐Encapsulated Black Pepper Oleoresin using Hydroxypropyl Beta‐Cyclodextrin for Antioxidant and Antimicrobial Applications

Previous studies have reported antimicrobial and antioxidant activity of black pepper oleoresin which is associated to its phenolic compounds and piperine. The ability of cyclodextrins to form an inclusion complex with a guest molecule could improve black pepper oleoresin application, bioavailability, and stability in foods. Hydroxypropyl beta‐cyclodextrin (HPBCD) inclusion complex with black pepper olereosin were synthesized using the kneading method and characterized for its physico‐chemical properties and its antioxidant and antimicrobial activities. Inclusion complex size was 103.9 ± 7.6 nm and indicated to be a polydisperse system. The entrapment efficiency was 78.3 ± 3.6%, which suggests that other constituents in black pepper oleoresin have higher affinities for HPBCD than piperine (major compound in black pepper oleoresin). Thermograms showed the disappearance of oxidation peaks of black pepper oleoresin, proving complex formation with HPBCD. Phase solubility results indicated 1:1 stoichiometric inclusion complex formation and an increase of black pepper oleoresin aqueous solubility with HPBCD concentration. Nano‐encapsulation with HPBCD did not affect (P > 0.05) total phenolic content; however, it enhanced (P < 0.05) black pepper oleoresin antioxidant activity. Black pepper oleoresin and its inclusion complex were analyzed for their antimicrobial activity against Escherichia coli K12 and Salmonella enterica serovar Typhimurium LT2. Both free and encapsulated black pepper oleoresin effectively inhibited bacterial growth within the concentration range tested. Black pepper oleoresin encapsulated in HPBCD was able to inhibit Salmonella at lower (P < 0.05) concentrations than its corresponding free extract. Therefore, black pepper oleoresin‐HPBCD nanocapsules could have important applications in the food industry as antimicrobial and antioxidant system.     

Purification and characterization of an amylase from Opuntiaficus‐indica seeds

BACKGROUND: In Tunisia, prickly pear fruit grow spontaneously; it is consumed as fresh fruit, juice or jam. When the fruit is used for juice production, the seeds are discarded and go to waste. Our study aimed to extract biomolecules from seeds by producing value‐added products from the fruits. RESULTS: An amylase from Opuntia ficus‐indica seeds was extracted and purified to homogeneity. An increase in specific activity of 113‐fold was observed. The apparent molecular mass of the enzyme is 64 kDa. The optimum pH and temperature for enzyme activity were pH 5 and 60 °C, respectively. Under these conditions, the specific activity is 245.5 U mg^?1. The enzyme was activated by Co²⁺ and Mg²⁺ (relative activity 117% and 113% respectively) at lower ion concentrations. It was strongly inhibited by Mn²⁺ and Fe²⁺. Cu²⁺ inhibited totally the activity of this enzyme, but Ca²⁺ has an inhibitory effect which increases with ion concentration. CONCLUSION: The extracted enzyme belongs to the exo type of amylases and is classified as a β‐cyclodextrin glycosyltransferase since it generates mainly β‐cyclodextrin from starch. It exhibits high thermal stability and a broad range of pH stability, making it a promising prospect for industrial and food applications. Copyright © 2012 Society of Chemical Industry     

Production,Characterization, and Stability of Orange or Eucalyptus Essential Oil/β‐Cyclodextrin Inclusion Complex

The aim of this study was to produce and characterize inclusion complexes (IC) between β‐cyclodextrin (β‐CD) and orange essential oil (OEO) or eucalyptus essential oil (EEO), and to compare these with their pure compounds and physical mixtures. The samples were evaluated by chemical composition, morphology, thermal stability, and volatile compounds by static headspace‐gas chromatography (SH‐GC). Comparing the free essential oil and physical mixture with the inclusion complex, of both essential oils (OEO and EEO), it was observed differences occurred in the chemical composition, thermal stability, and morphology. These differences show that there was the formation of the inclusion complex and demonstrate the necessity of the precipitation method used to guarantee the interaction between β‐CD and essential oils. The slow loss of the volatile compounds from both essential oils, when complexed with β‐CD, showed a higher stability when compared with their physical mixtures and free essential oils. Therefore, the results showed that the chemical composition, molecular size, and structure of the essential oils influence the characteristics of the inclusion complexes. The application of the β‐CD in the formation of inclusion complexes with essential oils can expand the potential applications in foods.     

Extraction of Epigallocatechin Gallate and Epicatechin Gallate from Tea Leaves Using β‐Cyclodextrin

Use of organic solvents to extract phenolic compounds from plants may result in environmental pollution and cause health problems in persons. Replacing organic extraction solvents by green extracting agents without affecting the extraction yield is one of the most pressing problems to be solved. The aim of this study is to evaluate the capacity of β‐cyclodextrin (β‐CD) to recover phenolic compounds from tea leaves. The extract obtained using the ethanol/water mixture presented the highest total phenolic content, followed by those obtained using β‐CD solution and water. HPLC analysis of the extracts showed that the addition of β‐CD to the extracting agent had a selective effect on the extraction of epigallocatechin gallate (EGCG) and epicatechin gallate (ECG). The extraction yield of EGCG and ECG using 15 g/L β‐CD were higher than that obtained using water and 50% ethanol. Molecular docking results indicated that the molecules of EGCG and ECG were more inclined to interact with β‐CD than epigallocatechin, epicatechin, and gallocatechin. The impact of β‐CD concentration, temperature, and time on EGCG and ECG extraction from tea leaves was investigated and the maximum amount of EGCG (118.7 mg/g) and ECG (54.6 mg/g) were achieved when extracted with 25 g/L aqueous β‐CD solution at 60 °C for 60 min. The present study indicates that aqueous β‐CD can be used as an alternative to organic solvents to recover EGCG and ECG from tea leaves.     

Characteristics of Modified β‐Cyclodextrin Bound to Cellulose Powder

Monochlorotriazinyl‐β‐cyclodextrin (MCT‐β‐CD) was covalently bonded to cellulose powder. The amount of MCT‐β‐CD bonded to cellulose could be determined by infrared spectroscopy. The coupling reaction was characterized as a physical adsorption of the MCT‐β‐CD on the cellulose powder followed by an apparently zero order chemical reaction. The reaction rate constant was found to be k = 6.43 · 10^‐4 ± 0.11 · 10^‐4 g g^‐1 s^‐1. The immobilized cyclodextrin was able to include and release d‐limonene as a model flavor compound. The maximum molar inclusion ratio was 0.85, which is the same inclusion ratio as for d‐limonene in native β‐cyclodextrin. The release rates of dlimonene included in the fixed MCT‐β‐CD were monitored at various relative humidities and 50 °C. The release kinetics could be modeled using the Avrami equation. These results demonstrate that flavors as well as other hydrophobic compounds can be included and released from MCT‐β‐CD immobilized on cellulose.     

Cyclodextrin and methacrylic acid octyl phenols poly(ethylene oxide) ester‐induced synergistic effect in a novel poly(AM‐co‐A‐β‐CD‐co‐AE) polymer

Owing to superior properties such as temperature resistance and salt tolerance etc., modified polyacrylamide (PAM) as one of the main injected polymers has been widely investigated to enhance oil production in reservoirs. Herein, a novel poly(AM‐co‐A‐β‐CD‐co‐AE) polymer was synthesized by utilizing β‐CD and AE to copolymerize with AM and characterized by FT‐IR and SEM. Furthermore, the temperature resistance and salt tolerance of poly(AM‐co‐A‐β‐CD‐co‐AE) polymer were explored. The results showed that the presence of the poly(AM‐co‐A‐β‐CD‐co‐AE) polymer better achieved temperature resistance and salt tolerance properties than is the case with PAM, which has potential application for enhancing oil recovery in the high‐temperature and high‐mineralization oilfield. On the other hand, the inhibition performance of poly(AM‐co‐A‐β‐CD‐co‐AE) polymer as a corrosion inhibitor was evaluated by SEM and electrochemical techniques. SEM observations of the carbon steel surface confirmed the protective role of the corrosion inhibitor. The results of potentiodynamic polarization and EIS measurements on the corrosion inhibition of carbon steel samples in 0.5 M sulfuric acid solutions revealed that the highest inhibition efficiency of it over 90% was obtained, indicating poly(AM‐co‐A‐β‐CD‐co‐AE) polymer acts as a more efficient corrosion inhibitor for carbon steel.     

The role of pulsed electric fields treatment in enhancing the stability of amino acid – sugar complexes:‐ interactions between L‐Phenylalanine and β‐Cyclodextrin

In this work, the effect of pulsed electric fields (PEF) treatment on the interactions between amino acids (using L‐Phenylalanine: L‐Phe) and sugar (using β‐Cyclodextrin: β‐CD) complex was analysed by fluorescence spectroscopy, Raman spectroscopy and simultaneous thermal analyzer. Moreover, the molecular dynamics of β‐CD–L‐Phe inclusion complex treated by PEF was calculated by molecular modelling. The results indicated that β‐CD–L‐Phe complexes are formed by a molar ratio of 1:1, and the stability constant of such complexes increased from 147 to 614 M^?1 by PEF treatment. Thermal characterisations of β‐CD–L‐Phe complexes indicated that the PEF treatment could increase the yield of complexes. The PEF treatment resulted in an increase in the reaction enthalpy of β‐CD–L‐Phe inclusion complexes by DSC curve. These results show that PEF treatment has the potential to promote the chemical processing, especially the small organic molecules participate in inclusion or cross‐linking reaction.     

Lactic acid‐mediated isolation of alpha‐, beta‐ and kappa‐casein fractions by isoelectric precipitation coupled with cold extraction from defatted cow milk

The present study focused on the isolation of individual caseins (α, β and κ) from defatted cow's milk using lactic acid‐mediated isoelectric precipitation coupled with cold extraction technique, followed by purification using sucrose‐based density gradient centrifugation method. In vitro characterisation of the isolated casein fractions confirmed preservation of micellar morphology throughout the experimental conditions with anionic surface charge and wider particle size range. The RP‐HPLC analysis showed a peak at retention time of 25.13 min and 27.84 min for α‐casein and β‐casein, respectively. Results confirmed isolation of pure casein fractions from cow's milk which could serve as a rich source of bioactive peptides for nutraceutical/pharmaceutical applications.     

Production of isomalto‐oligosaccharide syrup from rice starch using an one‐step conversion method

Isomalto‐oligosaccharides (IMO) belong to a group of prebiotics that can significantly increase the number of protective gut microflora. A one‐step method using neopullulanase (NPN) in conjunction with saccharifying α‐amylase (SAA) for the bioconversion of rice starch into IMO was investigated. Purified rice starch slurry (30% w/w) was mixed with NPN (3.5 U g^?1 starch substrate) and SAA (6.5 U g^?1 starch substrate) and the slurry was incubated at 57 °C for 92 h under constant stirring. The carbohydrate composition of the resulting syrup was analysed by high performance liquid chromatography (HPLC) and the dextrose equivalent (DE) determined by titration. The amount of IMO in the syrup reached maximum (59.2%, dry basis) after 72 h of bioconversion. The concentration of glucose and maltose, which were the main carbohydrate residues of the IMO syrup, was 25.5% while the concentration of other oligosaccharide residues was about 1.0%. The results demonstrate that rice starch is a suitable matrix for producing IMO syrup and the one‐step conversion procedure appears to be an efficient method for converting starch into IMO syrup.     

Encapsulation of Beta‐carotene in Lipid Microparticles Stabilized with Hydrolyzed Soy Protein Isolate: Production Parameters,Alpha‐tocopherol Coencapsulation and Stability Under Stress Conditions

The objectives of this research were to study the encapsulation of beta‐carotene (BC) in solid lipid microparticles (SLM) of palm stearin (PS) and stabilized with hydrolyzed soy protein isolate (HSPI), and also to investigate the effect of alpha‐tocopherol (TOC) addition to the systems. Through the characterizations of SLM produced with different formulations, it was verified that systems with 5% (w/v) PS, 1.0% (w/v) HSPI, and 0.3% (w/v) xanthan gum (XG) presented the highest stability, with average diameters of approximately 1.2 μm. This formulation was applied for the production of BC‐loaded SLM, with different concentrations of TOC. In SLM containing TOC, nearly 75% of encapsulated BC was preserved after 45 d of storage. The kinetic profiles for degradation of encapsulated BC were fitted to a pseudo‐1st‐order model, and the results showed that the main difference among the systems with different BC:TOC ratios was the residual concentration of BC. The stability of the BC‐loaded SLMs was also studied after stress conditions, and the results showed that the SLMs were able to support thermal treatments over 60 °C but presented low stability after different ionic strength stresses.