利用农业废弃物碳源的红法夫酵母生产虾青素研究进展

利用红法夫酵母产虾青素工艺具有绿色、安全、低成本等优点,但培养红法夫酵母的培养基(尤其是碳源)成本是制约该工艺实现大规模生产的主要因素之一。传统的利用纯糖如葡萄糖、蔗糖、木糖等作为红法夫酵母生产虾青素的碳源,效果虽好,但因其价格较高,仅适用于实验室研究和小型发酵生产,不适于大规模工业化生产。如何寻找来源广、价廉且发酵效果好的碳源,降低虾青素的生产成本是发展虾青素产业亟待解决的课题。利用廉价且来源广泛的原料如水果类原料、糖质原料、甘油及废弃木质纤维素类原料等作为碳源能有效降低成本。但这些原料都存在一些问题,比如水果类原料存在季节性及地域性限制,糖蜜类原料供应稳定性方面存在问题,甘油类原料杂质多且存在食品安全隐患,木质纤维素类原料来源广泛、价格低廉,不存在占用耕地等问题,是最有希望用于规模化低成本发酵生产虾青素的原料,但其利用存在一些技术瓶颈。该文介绍了目前利用红法夫酵母生产虾青素的基本情况,并系统地对国内外不同碳源条件下利用红法夫酵母生产虾青素的研究状况进行了综述、比较及分析。最后,对利用红法夫酵母生产虾青素在碳源方面的未来发展趋势及重点研究方向进行了展望。     

超临界CO2萃取南美白对虾虾青素的工艺优化

为提高虾青素萃取物得率和虾青素纯度,采用超临界CO2萃取南美白对虾虾头废弃物中的虾青素,皂化后用高效液相色谱对虾青素含量进行测定。单因素试验确定萃取参数对虾青素萃取物得率和虾青素纯度的影响,然后进一步运用响应面法对萃取工艺参数进行优化。结果表明：超临界CO2萃取工艺参数对虾青素萃取物得率影响不显著（P＞0.05）,但对虾青素纯度影响显著（P＜0.05）;物料粒径40目,质量含水率9%时,响应面理论优化最佳萃取参数为萃取压力403.95 Pa,萃取温度39.95℃,CO2流量1.16 L/min,虾青素纯度为796.3 mg/g。结合实际应用,验证试验在萃取压力400 Pa,萃取温度40℃,流量1.2 L/min时,虾青素纯度为789.61 mg/g。研究结果可为虾青素的提取和纯化提供参考。     

Preparation of ferulic acid from agricultural wastes: its improved extraction and purification

Ferulic acid (FA) is a phenolic antioxidant present in plants, which is widely used in the food and cosmetic industry. In the present study, various agricultural wastes such as maize bran, rice bran, wheat bran, wheat straw, sugar cane baggasse, pineapple peels, orange peels, and pomegranate peels were screened for the presence of esterified FA (EFA). Among the sources screened, maize bran was found to contain the highest amount of EFA. Pineapple peels, orange peels, and pomegranate peels were also found to contain traces of EFA. Alkaline extraction of EFA from maize bran was carried out using 2 M NaOH. Response surface methodology (RSM) was used for optimization of EFA extraction, which resulted in a 1.3-fold increase as compared to the unoptimized conventional extraction technique. FA was analyzed by means of high-performance liquid chromatography (HPLC). Purification was carried out by adsorption chromatography using Amberlite XAD-16 followed by preparative high-performance thin-layer chromatography (HPTLC). The recovery of Amberlite XAD-16 purified FA was up to 57.97% with HPLC purity 50.89%. The fold purity achieved was 1.35. After preparative HPTLC, the maximum HPLC purity obtained was 95.35% along with an increase in fold purity up to 2.53.     

果蔬垃圾重复批次发酵协同制备乳酸和短链脂肪酸

利用果蔬垃圾厌氧合成中链脂肪酸（medium-chain fatty acids,MCFAs）等化学品是厌氧技术高值化的重要方向。中链脂肪酸合成通常需要以乳酸/乙醇（电子供体）和短链脂肪酸（电子受体）为碳源进行碳链延长反应。因此,利用有机废弃物连续、稳定地协同制备乳酸和短链脂肪酸是中链脂肪酸合成的关键步骤。该研究考察了果蔬垃圾重复批次发酵协同制备乳酸和短链脂肪酸（short-chain fatty acids,SCFAs）的可行性,研究了不同置换率和进料浓度对果蔬垃圾重复批次发酵产酸特性的影响。结果表明,调控重复批次发酵的置换率和进料浓度是提高生产率、改善乳酸/SCFAs比例的有效方法。综合考虑酸化产物的生产率、乳酸/SCFAs比例和碳源浓度,在70%置换率和8%进料TS(total solid)浓度条件下获得的酸化产物相对更有利于MCFAs的合成。此时,酸化产物生产率达到（5.25±0.25）g/(L·d),乳酸/SCFAs的碳摩尔比例达到5±0.3,碳源浓度达到（985±29）mmol C/L。微生物群落分析显示,乳酸菌,如Lactobacillus和Enterococcus作为优势菌通过异型乳酸发酵协同制备乳酸和SCFAs。研究结果可为果蔬垃圾的高值化利用提供参考。     

龙眼果浆复合乳酸菌发酵工艺优化

为筛选出适合龙眼发酵的复合乳酸菌,优化建立其发酵工艺,比较分析了7种不同的乳酸菌发酵龙眼的降糖能力和释放结合态酚类物质的能力,发现明串珠菌降糖能力最强,植物乳杆菌释放结合态酚能力最强。利用植物乳杆菌和明串珠菌复合发酵龙眼果肉,以游离酚质量分数和还原糖质量分数结合的综合模糊评判值为指标,通过Box-benhnken中心组合试验设计,确定复合乳酸菌发酵龙眼的最佳工艺条件。结果表明,各因素对复合乳酸菌发酵龙眼果浆的模糊评判值影响程度从强到弱依次为:发酵时间发酵温度接种量料液比。复合乳酸菌发酵龙眼的最佳工艺为:料液比为1∶7(g/m L),发酵温度为37℃,发酵时间为53 h,接种量为1.4%,菌种配比为1∶1。在最佳工艺条件下,发酵龙眼果浆的游离酚质量分数为(161.77±3.93)mg/(100 g),还原糖质量分数为(122.80±3.64)mg/g。该研究为开发低糖含量,且适合糖尿病、肥胖症等病人食用的龙眼乳酸菌发酵饮料新产品提供参考。     

产脂肪酶乳酸菌对羊肉发酵香肠脂肪酸的影响

脂肪酶在肉制品加工中会影响风味物质的积累与产生,发酵肉制品生产中常通过添加产脂肪酶菌株来改善制品的风味。通过乳酸菌在代谢过程中分解三丁酸甘油酯产生透明圈的大小,判断菌株产脂肪酶的活力,同时结合脂肪酶基因Lip0069、Lip0893表达量的变化情况,筛选产脂肪酶能力较高的乳酸菌,研究该菌株对发酵香肠中脂肪酸的影响。结果表明,瑞士乳杆菌TR1-1-3代谢过程中产酶活力最强,编码脂肪酶的基因Lip0069、Lip0893表达量最高。发酵香肠试验组共检测到34种脂肪酸,其中饱和脂肪酸（Saturated Fatty Acids, SFA）16种、单不饱和脂肪酸酸（Monounsaturated Fatty Acids, MUFA）9种和多不饱和脂肪酸（Polyunsaturated FattyAcids, PUFA）9种。含量较高的MUFA主要是油酸（C18:1C9）、棕榈油酸（C16:1）、十七碳烯酸（C17:1）、肉豆蔻烯酸（C14:1）。含量较高的PUFA主要是亚油酸（C18:2C9）、亚麻酸（C18:3N6、C18:3N3）、花生四烯酸（C20:4）、二十二碳六烯酸（C22:6,Docosahexaenoic Acid,DHA）。TR1-1-3和ZF22组发酵2 d后,MUFA的含量明显上升（P<0.05）。12 d时5个试验组MUFA的含量均达到最高,其中TR1-1-3增长比例最高,且与ZR、M组差异显著（P<0.05）。TR1-1-3组发酵2 d后PUFA的含量发生了明显的变化（P<0.05）,12 d时TR1-1-3组 PUFA的含量增长了54.17%,且与其他试验组相比差异显著（P<0.05）。乳酸菌TR1-1-3可降低产品中SFA在脂肪酸中所占比例,明显加快MUFA、PUFA的增加速度及其含量,可为研究改善香肠脂肪特性提供良好的前景。     

An efficient method for extraction of astaxanthin from green alga Haematococcus pluvialis

Haematococcus pluvialis is one of the potent organisms for production of astaxanthin, a high value ketocarotenoid. Astaxanthin is accumulated in thick-walled cyst cells of Haematococcus. The thick cell wall is made up of sporopollenin-like material, algaenan, which hinders solvent extraction of astaxanthin. In the present study, an improved method for extraction of astaxanthin without homogenization of cells is reported. Extractability of astaxanthin from cyst cells was evaluated by treating cells with various solvents and pretreating the cells with organic and mineral acids at 70 degrees C followed by acetone extraction. Hydrochloric acid treatment facilitated 86-94% extractability of astaxanthin. Treatment time, temperature, and concentration of the acid were found to be critical factors for maximum extractability. The treatment did not affect the astaxanthin ester profile and the treated cells can be preserved until further use.     

Antiallergic effect of milk fermented with lactic acid bacteria in a murine animal model

The objective of this study was to assess the antiallergic effect of fermented milk prepared, respectively, with Streptococcus thermophilus MC, Lactobacillus acidophilus B, Lactobacillus bulgaricus Lb, L. bulgaricus 448, and Bifidobacterium longum B6. Female BALB/c mice fed fermented milk were immunized intraperitoneally with ovalbumin (OVA)/complete Freund's adjuvant (CFA) to evaluate the immune response by observing the secretion of cytokines IL-2, IL-4, and IFN-gamma and serum antibody IgE. The results showed that supplementation with lactic acid bacteria fermented milk did not significantly change the IL-2 spontaneous and OVA-stimulated secretions of splenocytes. However, both spontaneous and OVA-stimulated secretions of splenocytes from mice fed lactic acid bacteria fermented milk showed significantly (P < 0.05) lower levels of IL-4 (Th2 cytokine) than those from OVA/CFA-immunized mice fed non-fermented milk (OVA/CFA-milk group). The spontaneous secretion of IFN-gamma (Th1 cytokine) by splenocytes from mice fed L. bulgaricus 448 or L. bulgaricus Lb fermented milk significantly increased as compared to that from the OVA/CFA-milk group. The results showed that the ratios of IFN-gamma to IL-4 of both spontaneous and OVA-stimulated secretions in splenocytes from mice fed lactic acid bacteria fermented milk increased significantly as compared to that of PBS- or OVA/CFA-milk groups. The serum levels of OVA-specific IgE in fermented milk fed groups, especially the group fed S. thermophilus MC fermented milk, were significantly lower than those in the OVA/CFA-milk group through a 6 week feeding experiment. The results showed that milk fermented with lactic acid bacteria demonstrated in vivo antiallergic effects on OVA/CFA-immunized mice via increasing the secretion ratio of IFN-gamma/IL-4 (Th1/Th2) by splenocytes and decreasing the serum level of OVA-specific IgE.     

Novel fibrinolytic enzyme in fermented shrimp paste, a traditional asian fermented seasoning

A novel fibrinolytic enzyme was purified from fermented shrimp paste, a popular seasoning used in Asian countries. The enzyme is a monomer with an apparent molecular weight of 18 kDa, and it is composed primarily of beta-sheet and random coils. The N-terminal amino acid sequence was determined to be DPYEEPGPCENLQVA. It is a neutral protease with an optimal activity from pH 3 to 7. No inhibition was observed with PMSF, Pepstatin A, E64, and 1,10-phenanthroline, but the enzyme was slightly inhibited by EDTA and Cu(2+). It was relatively specific to fibrin or fibrinogen as a protein substrate, yet it hydrolyzed none of the plasma proteins in the studies. In vitro, the enzyme was resistant to pepsin and trypsin digestion. It also had an anticoagulant activity measured with activated partial thrombin time and prothrombin time tests. The novel fibrinolytic enzyme derived from traditional Asian foods is useful for thrombolytic therapy. In addition, this enzyme has a significant potential for food fortification and nutraceutical applications, such that its use could effectively prevent cardiovascular diseases.     

Extraction, characterization, and nematicidal activity of chitin and chitosan derived from shrimp shell wastes

Chitin has been chemically extracted from the Egyptian shrimp shell waste. The obtained chitin was transformed into the more soluble chitosan. Chitin and chitosan were characterized using Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectrometry. These products were also characterized by their nematicidal potential against the root-knot nematode, Meloidogyne incognita, infecting tomato in a glasshouse. Chitin or chitosan was incorporated into the soil at the rate of 1, 3, 5, and 10 g/kg, and their nematicidal activity was compared with that of the synthetic nematicide oxamyl at the rate of 0.01 g a.i./kg. The effects of the treatments on the growth of tomato were also examined. The obtained results show that chitin and chitosan reduced tomato root galls and J₂ of M. incognita in the soil in a dose-dependent manner. Chitosan was more effective in the reduction of galls and J₂ in the soil than chitin. However, the efficacy of chitin and chitosan in reducing the number of J₂ in the soil was less than that of oxamyl. As compared to the untreated inoculated plants, the length of shoots and roots was significantly (p ≤ 0.05) increased in soil amended with either chitin or chitosan, whereas both treatments did not significantly (p > 0.05) differ with respect to the inoculated control on the effect on the dry weight of shoot and root systems. Furthermore, neither chitin nor chitosan treatments at the tested rates were phytotoxic to tomato plants.     

酸法破壁条件对法夫酵母虾青素提取效果的影响

为了评价酸法破壁在法夫酵母虾青素提取中的应用价值,从酸的种类、酸的浓度、液料比、破壁温度及时间等方面研究了条件因子对破壁后法夫酵母虾青素提取的影响并对破壁条件进行了优化。试验结果表明：乳酸比盐酸和醋酸更适合用于法夫酵母的破壁处理;酸的浓度、破壁温度等因素对虾青素的提取得率影响较大,而酸处理时间和液料比对虾青素的提取得率影响不大。用正交试验方法得到优化的乳酸破壁法夫酵母的条件为：乳酸浓度6 mol/L、破壁温度30℃、破壁时间5 min、酸加量15 mL/g(干菌体),优化后虾青素的提取得率为1374.7 μg/g(干菌体),总类胡萝卜素的提取得率为1709.9 μg/g(干菌体),提取物中虾青素占总类胡萝卜素80.4%。优化的破壁条件温和,不会引起残留毒性,能获得良好的虾青素提取效果,具有较好的工业应用前景。     

应用高电压脉冲电场辅助快速提取虾壳壳聚糖

为提高虾壳的综合利用效果,该文以虾壳为原料,将高电压脉冲电场（PEF）技术用于壳聚糖的提取研究。与常规加热法和微波法进行了对比试验,并通过单因素试验和三元二次回归旋转组合设计确定最佳制备工艺。结果表明：该技术与传统加热和微波法方法相比,具有非热、反应速度快,脱乙酰度高的特点。建立了脉冲数、场强、NaOH质量浓度对脱乙酰度影响的数学模型,确定了最佳工艺参数：脉冲数10,电场强度20.48 kV/cm,NaOH质量分数48.64%,该条件下脱乙酰度达到最大92.32%。因此,利用高电压脉冲电场技术可以快速制备虾壳中的壳聚糖。     

Characterization of high purity lycopene from tomato wastes using a new pressurized extraction approach

In this paper, a method for the extraction of high purity lycopene from tomato wastes is presented. The method is based on a pressurized extraction that uses the Extractor Naviglio, and it is performed in the 0.7-0.9 MPa range. Tomato skin, the byproduct deriving from manufacturing of tomato, in a water dispersion, are used as starting material. Lycopene is transferred, for the effect of the high pressure used, in the form of molecular aggregates into the water as a dispersion, while apolar compounds remain in the matrix. The aggregates are easily purified in a single subsequent step by using methanol, thus, obtaining lycopene at 98% chromatographic purity or higher. A new stationary phase, phenyl-hexyl silicone, and a simple water/acetonitrile gradient were used for HPLC analysis of lycopene. The extract was characterized by UV-Vis spectrophotometry, (1)H NMR, (13)C NMR, and electrospray ionization mass spectrometry. An average recovery of 2.8 mg lycopene/kg tomato waste can be obtained after 4 hours of extraction and using tap water as the extracting liquid. The recovery percentage was of about 10%. The exhausted tomato byproduct can be easily dried and used in agriculture or as feeding for animals.     

辐照对发酵玉米芯中裂褶菌多糖提取及流变学性质的影响

对裂褶菌发酵玉米芯进行γ射线辐照处理,研究不同处理条件下辐照对裂褶菌多糖提取率及流变学性质的影响。结果显示:在5~20kGy辐照剂量范围内,同一剂量下,多糖提取率随辐照原料水分含量的增加而上升,60%水分含量、10kGy处理下多糖提取率达到最高值7.95%。辐照处理对所提取裂褶菌多糖溶液粘度有不同程度的降低,15.0%水分含量、20kGy处理下,多糖的零剪切粘度η0降至最低值10.79 Pa.s;辐照处理对裂褶菌多糖的粘弹性亦有显著影响,G′、G″以及二者之交点Gx值的变化趋势与η0值的基本一致。本研究得到如下结论:γ射线辐照处理对发酵玉米芯中裂褶菌多糖可起到降解、增溶的作用。     

Antioxidative ability of lactic acid bacteria

Nineteen strains of lactic acid bacteria were investigated for antioxidative activity. These includedLactobacillus acidophilus B, E, N1, 4356, LA-1, and Farr; Lactobacillus bulgaricus 12 278, 448, 449, Lb, 1006, and 11 842; Streptococcus thermophilus 821, MC, 573, 3641, and 19 987; and Bifidobacterium longum B6 and 15 708. Intracellular cell-free extract of all strains demonstrated antioxidative activity with inhibition rates of ascorbate autoxidation in the range of 7-12%. Antioxidative mechanisms including metal ion chelating ability, scavenge of reactive oxygen species, enzyme inhibition, and reducing activity of intracellular cell-free extract of lactic acid bacteria were studied. S. thermophilus 821 had the highest metal ion chelating ability for Fe(2+), and B. longum 15 708 showed the highest Cu(2+) chelating ability among the 19 strains tested. All strains demonstrated reactive oxygen species scavenging ability. L. acidophilus E showed the highest hydroxyl radical scavenging ability, and B. longum B6 had the best hydrogen peroxide scavenging ability. Reducing activity was also found in all strains. Most of the strains tested demonstrated excellent reducing activity. B. longum B6 showed the highest reducing activity among the 19 strains tested. In enzyme inhibition, superoxide dismutase activity was not found in these 19 strains, and the activity of superoxide dismutase was not induced when metal ion Mn(2+), Fe(2+), or Cu(2+)Zn(2+) was present.     

菜籽饼粕中多糖的酸提取工艺优化

采用五因子二次正交旋转设计,研究酸浓度、料液比、提取时间、提取温度、提取次数等5个因素对酸提取菜籽饼粕中多糖的得率的影响,用SAS8.0中响应面分析程序RSREG建立并分析了5个因素对得率的函数模型。结果表明,所得回归方程达显著水平,拟合情况良好;5个因素对得率的影响大小依次为提取温度>提取次数>提取时间>酸浓度>料液比,最佳提取温度为100℃,提取次数4～5次,提取时间(每次)1.4～1.8 h,酸浓度0.10～0.14 mol/L,料液比1∶18～1∶22,菜籽饼粕多糖得率在3.1%以上。     

PESA对污泥中铬的萃取研究

An environmentally benign biodegradable chelant,polyepoxysuccinic acid(PESA),was used to separate heavy metals from sewage sludge from the Shanghai Taopu Wastewater Treatment Plant,China,based on chemical extraction technology.The extraction of chromium(Cr) from sewage sludge with an aqueous solution of PESA was studied under various conditions.It was found that the extraction of Cr using PESA was more efficient than that using ethylenediaminetetraacetic acid(EDTA) and S,S-ethylenediaminedisuccinic acid(EDDS) under similar conditions.PESA was capable of extracting Cr from the sewage sludge,and the extraction efficiency was obviously dependent on both the pH and the concentration of the chelating reagent.The extraction efficiency decreased gradually with increasing pH,and the dependence on pH decreased as the concentration of PESA increased.The extraction efficiency reached 58% under conditions of pH = 4 and a ratio of PESA to total heavy metals of 10:1.The extraction efficiency was maintained above 40% within the pH range from 1 to 7 at the high ratio of PESA to total heavy metals of 10:1.Comparing the contents of heavy metals in the sewage sludge before and after the extraction,it was found that the extracted Cr came mainly from the reducible and oxidizable fractions.     

Effects of acid mine wastes on aquatic ecosystems

The Cedar Creek Basin (39th N parallel 92nd W meridian) was studied for the period June 1952 through August 1954 to observe the effects of both continuous and periodic acid effluent flows on aquatic communities. The acid strip-mine effluent contained ferric and ferrous Fe, Cu, Pb, Zn, Al, Mg, titratable acid, and elevated H ion concentration, and was toxic to many of the aquatic organisms. In the areas of Cedar Creek, subjected to continuous acid flow, planktonic and benthic species had become adapted to the severe conditions and varied in abundance and diversity. No fishes were observed in the continuous acid effluents area. Downstream, where periodic mineral acid conditions changed drastically during an excessive effluent flow, planktonic and benthic communities had high diversity but low density. The populations of fishes were variable in this stream reach. The chemical basis of water quality variability was shown to be time-related, and statistically related to the aquatic communities. Physical, chemical and biological conditions of acid lakes formed by surface mining were reviewed. In addition to apparent physical and chemical differences in lakes, due in part to variable solar absorbance due to suspended oxides of Fe, chemical variability related to organic composition was reviewed. It is quite probable that degradation of such lakes can be reversed.     

Isolation and characterization of lactic acid bacteria from feces of newborn baby and from dongchimi

Lactic acid bacteria were screened from feces of newborn baby and from dongchimi. Selection criteria employed included the ability of strains to withstand environmental conditions such as low pH, high bile concentration, and oxygen. The isolates were applied to the juice of various vegetables, and fermentabilities of isolates were compared. Strains F20-3, F35-3, and F35-6 showed high stability compared to the other strains at pH 3.0 and 2.3. Strains D1 and D2 showed the highest survival at pH 3.0 and survived at 1% high bile concentration. The selected strains were able to survive at low pH and relatively high bile concentration and were not affected by oxygen. The growth of isolates was >10(7) cfu/mL in natural media, and strains were not affected by the pH values of the vegetables. Therefore, isolated strains are thought to survive through the intestinal ecosystem and are considered to be suitable for application of the fermented product using various vegetables for their functionality. The isolates were identified as Lactobacillus plantarum and Lactobacillus fermentum.