以菊芋粉为原料同步糖化发酵生产燃料乙醇

利用粟酒裂殖酵母（Schizosaccharomyces pombe）能发酵菊芋未水解糖液高产乙醇的特点提出了以菊芋粉为原料,同步糖化发酵生产燃料乙醇的新工艺。在摇瓶中考察了原料预处理方法、原料浓度和初始pH值对乙醇发酵的影响,进而在5 L发酵罐中考察了未调控pH值和恒定pH值与通气情况对乙醇发酵的影响。结果表明：该菌株最适pH值为4.0;100目筛分的菊芋粉发酵效果良好,115℃灭菌处理优于121℃,在此条件下,菊芋粉浓度200 g/L时,乙醇产量达到66.58 g/L,理论转化率为85.88%;发酵液pH值下降对乙醇发酵没有影响,通入适量氧气会导致乙醇产量的下降,这表明粟酒裂殖酵母进行乙醇发酵时不需要供氧;通入氮气保持厌氧环境不能显著提高乙醇产量,不通气进行乙醇发酵也达到高的转化率,因此在工业生产中,不必保持厌氧发酵环境。在此基础上,对菊芋粉补料发酵进行了试验,补料至菊芋粉终浓度为300 g/L,发酵终点乙醇浓度为94.81 g/L,理论转化率为81.54%。这些研究工作,为以菊芋为原料的燃料乙醇工业化生产提供技术依据。     

玉米秸秆湿氧化预处理同步糖化发酵酒精

为了找到适合的玉米秸秆生产酒精工艺,该文采用碱性湿氧化预处理（195℃,15 min,Na2CO3 2 g/L,O2 1 200 kPa）与同步糖化发酵对玉米秸秆制备酒精进行了研究。结果表明：经过预处理,90%纤维素保留在固体中,回收率为95.87%。固体部分利用纤维素酶处理,在50℃ 24 h酶解率达到了67.6%。底物8%（质量分数）,经过142 h同步糖化发酵,酒精产量达到了理论酒精产量的79.0%。假定五碳糖和六碳糖都能够被利用,相当于1 t玉米秸秆能够产生262.7 kg的酒精。发酵过程中没有明显的抑制作用。该文为玉米秸秆发酵生产酒精提供了试验数据。     

汽爆玉米秸秆同步糖化发酵产乙醇的工艺优化

该文对同步糖化发酵玉米秸秆生产纤维乙醇的工艺条件进行了研究,分别利用单因素试验和正交试验考察了影响发酵的工艺条件。结果显示：温度37℃、pH值4.8、底物15%（质量分数）、表面活性剂Tween-20 0.15%（质量分数）、纤维素酶用量30 U/g、酵母用2%的葡萄糖溶液活化,接种量5‰（相对底物干质量）、培养基的组成：KH2PO4 2.5 g/L,(NH4)2SO4 2.0 g/L,MgSO4·7H2O 0.15 g/L,CaCl2 0.35 g/L、发酵时间60 h为同步糖化发酵（SSF）生产乙醇的最优工艺条件,该条件下乙醇的体积分数为2.85%,乙醇得率达理论值的88.12%。     

甜高粱茎汁及茎渣同步糖化发酵工艺优化

为了提高甜高粱秸秆乙醇生产中茎汁和茎渣的利用,以甜高粱茎汁及其渣为发酵原料,对茎汁茎渣混合原料同步糖化乙醇发酵的工艺条件进行优化研究。采用Plackett-Burman(PB)筛选设计试验筛选出影响甜高粱茎秆渣汁同步糖化乙醇发酵的显著因素。采用响应面法建立了同步糖化发酵乙醇生产的乙醇产量数学模型。根据该模型进行了工艺参数的优化,以乙醇产量为指标,试验所得甜高粱茎秆渣汁同步糖化化乙醇发酵的优化工艺条件为:发酵温度36.58℃,混合纤维素酶添加量=23.5(FBU/m L)/35.25(CBU/m L),甜高粱渣汁质量体积比为8.2%,理论预测乙醇产量为89.2%,在此条件下进行验证试验,乙醇产量为88.98%,平均质量浓度,验证了数学模型的有效性,为提高甜高粱茎汁及茎渣混合原料同步糖化发酵产乙醇和提高发酵效率提供参考。     

Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass

Simultaneous saccharification and fermentation (SSF) of alkaline hydrogen peroxide pretreated Antigonum leptopus (Linn) leaves to ethanol was optimized using cellulase from Trichoderma reesei QM-9414 (Celluclast from Novo) and Saccharomyces cerevisiae NRRL-Y-132 cells. Response surface methodology (RSM) and a three-level four-variable design were employed to evaluate the effects of SSF process variables such as cellulase concentration (20-100 FPU/g of substrate), substrate concentration (5-15% w/v), incubation time (24-72 h), and temperature (35-45 degrees C) on ethanol production efficiency. Cellulase and substrate concentrations were found to be the most significant variables. The optimum conditions arrived at are as follows: cellulase = 100 FPU/g of substrate, substrate = 15% (w/v), incubation time = 57.2 h, and temperature = 38.5 degrees C. At these conditions, the predicted ethanol yield was 3.02% (w/v) and the actual experimental value was 3.0% (w/v).     

Synthesis of structured triacylglycerols containing caproic acid by lipase-catalyzed acidolysis: optimization by response surface methodology.

Production in a batch reactor with a solvent-free system of structured triacylglycerols containing short-chain fatty acids by Lipozyme RM IM-catalyzed acidolysis between rapeseed oil and caproic acid was optimized using response surface methodology (RSM). Reaction time (t(r)), substrate ratio (S(r)), enzyme load (E(l), based on substrate), water content (W(c), based on enzyme), and reaction temperature (T(e)), the five most important parameters for the reaction, were chosen for the optimization. The range of each parameter was selected as follows: t(r) = 5-17 h; E(l) = 6-14 wt %; T(e) = 45-65 degrees C; S(r) = 2-6 mol/mol; and W(c) = 2-12 wt %. The biocatalyst was Lipozyme RM IM, in which Rhizomucor miehei lipase is immobilized on a resin. The incorporation of caproic acid into rapeseed oil was the main monitoring response. In addition, the contents of mono-incorporated structured triacylglycerols and di-incorporated structured triacylglycerols were also evaluated. The optimal reaction conditions for the incorporation of caproic acid and the content of di-incorporated structured triacylglycerols were as follows: t(r) = 17 h; S(r) = 5; E(l) = 14 wt %; W(c) = 10 wt %; T(e) = 65 degrees C. At these conditions, products with 55 mol % incorporation of caproic acid and 55 mol % di-incorporated structured triacylglycerols were obtained.     

Structural characterization of oligosaccharides and polysaccharides from apple juices produced by enzymatic pomace liquefaction

Eight apple pomace liquefaction juices were produced to characterize soluble cell wall material released by the action of pectolytic and cellulolytic enzyme preparations. Very high colloid values from 9.7 to 19.6 g/L were recovered from the juices by ethanol precipitation. The crude polysaccharides consisted mainly of galacturonic acid (49-64 mol %), arabinose (14-23 mol %), galactose (6-15 mol %), and minor amounts of rhamnose, xylose, and glucose. Separation of the polysaccharides by anion-exchange chromatography yielded one neutral, one slightly acidic, and one acidic polymer accounting for 60% of total colloids. Preparative size exclusion chromatography of the acidic fractions resulted in four polymers of different molecular weights and different sugar compositions. Among them, high molecular weight arabinans and rhamnogalacturonans as well as oligomeric fractions consisting of only galacturonic acid could be found. Linkage studies were performed on neutral fractions from anion-exchange chromatography and size exclusion chromatography. They revealed highly branched arabinans, xyloglucans, and mainly type I arabinogalactans.     

Optimization of lycopene extraction from tomato cell suspension culture by response surface methodology

Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold.     

Preparation of glucosamine from exoskeleton of shrimp and predicting production yield by response surface methodology

Chitin was prepared from Persian Gulf shrimp (Metapenaeus monoceros), and then, the obtained chitin was hydrolyzed by hydrochloric acid solutions. The production yield of glucosamine hydrochloride from chitin was optimized, and the effect of three factors (acid concentration, acid to chitin ratio, and reaction time) was investigated. A Box-Behnken design by Minitab software created 12 reactions with different conditions. Each reaction was performed in two replicates. Response surface methodology was used for predicting the glucosamine preparation. The optimum conditions for glucosamine hydrochloride preparation were 30 and 37% hydrochloric acid, 9:1 (v/w) acid solution to solid ratio, and 4 h of reaction time. Time ratio and time acid concentrations were the effective factors on the yield.     

Evaluation of the simultaneous effects of processing parameters on the iron and zinc solubility of infant sorghum porridge by response surface methodology

The purpose of this study was to improve the micronutrient quality of indigenous African infant flour using traditional techniques available in the region. Response surface methodology was used to study the effect of duration of soaking, germination, and fermentation on phytate and phenolic compounds (PC), pH, viscosity, and the in vitro solubility (IVS) of iron and zinc in infant sorghum flour. The phytate and the PC concentrations of the flour were significantly modified as a result of the duration of germination and fermentation and their mutual interaction. These modifications were accompanied by a significant increase in % IVS Zn after 24 h of sprouting. Except for the interaction of soaking and fermentation, none of the processing parameters exerted a significant effect on the % IVS Fe. The viscosity of the porridge prepared with the flour decreased significantly with the duration of germination, making it possible to produce a porridge with high energy and nutrient density. The use of germination in combination with fermentation is recommended in the processing of cereals for infant feeding in developing countries.     

Enzymatically catalyzed synthesis of low-calorie structured lipid in a solvent-free system: optimization by response surface methodology

A kind of low-calorie structured lipid (LCSL) was obtained by interesterification of tributyrin (TB) and methyl stearate (St-ME), catalyzed by a commercially immobilized 1,3-specific lipase, Lipozyme RM IM from Rhizomucor miehei . The condition optimization of the process was conducted by using response surface methodology (RSM). The optimal conditions for highest conversion of St-ME and lowest content LLL-TAG (SSS and SSP; S, stearic acid; P, palmitic acid) were determined to be a reaction time 6.52 h, a substrate molar ratio (St-ME:TB) of 1.77:1, and an enzyme amount of 10.34% at a reaction temperature of 65 °C; under these conditions, the actually measured conversion of St-ME and content of LLL-TAG were 78.47 and 4.89% respectively, in good agreement with predicted values. The target product under optimal conditions after short-range molecular distillation showed solid fat content (SFC) values similar to those of cocoa butter substitutes (CBS), cocoa butter equivalent (CBE), and cocoa butters (CB), indicating its application for inclusion with other fats as cocoa butter substitutes.     

Controlling molecular weight and degree of deacetylation of chitosan by response surface methodology

Response surface methodology (RSM) was used for controlling molecular weight (MW) and degree of deacetylation (DOD) of chitosan in chemical processing. In a reduced model, MW of chitosan is y = 1736166.406 - 250.745X(1)X(2) - 265.452X(2)X(3), with R( 2) = 0.86, and DOD of chitosan is y = 30.6069 + 0.3396X(1) + 0.4948X(2) + 0.0094X(3)(2), with R( 2) = 0.89. MW of chitosan depends on the crossproduct of temperature and NaOH concentration and the crossproduct of NaOH concentration and time, and DOD depends linearly on temperature and NaOH concentration, and quadratically on time. Chitosan was widely depolymerized in a range from 1,100 kDa to 100 kDa and deacetylated from 67.3 to 95.7% by NaOH alkaline treatment. MW and DOD of chitosan were drastically decreased and increased, respectively, with increase of temperature, reaction time, and NaOH concentration. Furthermore, the rate of MW decrease and DOD increase of chitosan gradually decreased with prolonged reaction time.     

响应面法优化盐渍藠头蒜素提取工艺参数

为了获得在自然pH值条件下利用果胶酶提取盐渍藠头中的风味物质蒜素的最佳工艺参数,以浸提温度、时间和果胶酶添加比为试验因子,蒜素含量为响应值,采用三元二次回归正交旋转组合设计进行了试验.结果表明,3个因素对蒜素含量的影响大小依次为提取温度＞保温时间＞果胶酶添加比;边际效应大小为提取温度＞保温时间＞果胶酶添加比;通过典形分析和岭脊分析确定果胶酶提取盐渍菖头中蒜素的最佳工艺参数为:浸提温度47℃、浸提时间3.3 h、果胶酶添加比0.77%,蒜素最高含量为0.274 g/(100 mL).所得回归模型拟合情况良好,达到设计要求.     

Optimization and modeling of glyphosate biodegradation by a novel Comamonas odontotermitis P2 through response surface methodology

Glyphosate is an important organophosphonate herbicide used to eliminate grasses and herbaceous plants in many vegetation management situations.Its extensive use is causing environmental pollution,and consequently,there is a need to remove it from the environment using an eco-friendly and cost-effective method.As a step to address this problem,a novel bacterial strain Comamonas odontotermitis P2,capable to utilize glyphosate as a carbon(C) and/or phosphorus(P) source,was isolated from a glyphostate-contaminated field soil in Australia and characterized.Response surface methodology(RSM)employing a 23 full factorial central composite design was used to optimize glyphosate degradation by C.odontotermitis P2 under various culture conditions.The strain C.odontotermitis P2 was proficient in degrading 1.5 g L~(-1) glyphosate completely within 104 h.The optimal conditions for the degradation of glyphosate were found to be pH 7.4,29.9℃,and an inoculum density of 0.54 g L~(-1),resulting in a maximum degradation of 90%.Sequencing of glyphosate oxidoreductase(GOX) and C-P lyase(phnJ) genes from C.odontotermitis P2 revealed 99% and 93% identities to already reported bacterial GOX and phnJ genes,respectively.The presence of these two genes in C.odontotermitis indicates its potential to degrade glyphosate through GOX and C-P lyase metabolic pathways.This study demonstrates the potential of C.odontotermitis P2 for efficient degradation of glyphosate,which can be exploited for remediation of glyphosate.     

凹土复配剂修复辣椒自毒作用的响应面优化及其机理研究

以邻苯二甲酸二丁酯(DBP)模拟辣椒连作自毒物质,选择凹土、牛粪、腐植酸为修复剂,采用BoxBehnken试验设计优化了复配参数,探讨了复配剂对辣椒DBP自毒作用的修复机理。结果显示,对DBP修复效果最好的复配剂为凹土20 g/kg+牛粪70 g/kg+腐植酸617 mg/kg,在此条件下,辣椒株高实测值与模型预测值相符。复配剂能显著增强辣椒根际土壤的脲酶、脱氢酶、蛋白酶和蔗糖酶活力,使辣椒根系受DBP毒害减轻,根系过氧化物酶(POD)和过氧化氢酶(CAT)活力增强,膜脂过氧化程度减弱,丙二醛(MDA)含量降低,根系活力提高。凹土复配剂对辣椒连作中DBP自毒作用有良好的修复作用。     

Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds

Dietary ellagic acid and related polyphenols are metabolized in humans to dibenzopyran-6-one derivatives, and the microbial origin of these metabolites has been suggested. However, this has not been demonstrated so far. Fecal samples donated by six volunteers were incubated under anaerobic conditions, and aliquots were used to evaluate the fecal metabolism of ellagic acid, the ellagitannin punicalagin, and an ellagitannin rich extract from walnuts. The isoflavone daidzein was also incubated with the same fecal samples to follow the production of the microbial metabolites previously reported (dihydrogenistein, O-demethylangolensin, and equol) as a positive control of the system and to evaluate similarities between isoflavone and ellagic acid fecal flora metabolism. After fermentation the metabolite "urolithin A" (3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one) was produced from ellagic acid, punicalagin, and the ellagitannin extract in all the fecal cultures from different volunteers, but with very different production rates and concentrations. This large variability in the concentration of metabolite and kinetics of metabolite production is consistent with the large variability found in the excretion of these metabolites in urine in vivo after human consumption of ellagitannins, and with differences in the composition of the fecal microflora. No correlation between isoflavone and ellagic acid metabolism by fecal microflora was observed. The present study confirms the microbial origin of the recently reported in vivo generated hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives in humans and is a further step in the study of the bioavailability and metabolism of ellagic acid and ellagitannins.     

Modeling of lipase-catalyzed acidolysis of sesame oil and caprylic Acid by response surface methodology: optimization of reaction conditions by considering both acyl incorporation and migration

Lipase-catalyzed acidolysis in hexane to produce structured lipids (SLs) from sesame oil and caprylic acid was optimized by considering both total incorporation (Y1) and acyl migration (Y2). Response surface methodology was applied to model Y1 and Y2, respectively, with three reaction parameters: temperature (X1), reaction time (X2), and substrate molar ratio (X3). Well-fitting models for Y1 and Y2 were established after regression analysis with backward elimination and verified by a chi2 test. All factors investigated positively affected Y1. For Y2, X1 showed the greatest positive effect. However, there was no effect of X3. We predicted the levels of Y2 and acyl incorporation into sn-1,3 positions (Y3) based on Y1. The results showed that over the range of ca. 55 mol % of Y1, Y3 started to decrease, and Y2 increased rapidly, suggesting that Y1 should be kept below ca. 55 mol % to prevent decrease in quality and yield of targeted SLs.     

Optimization of growth of Lactobacillus acidophilus FTCC 0291 and evaluation of growth characteristics in soy whey medium: a response surface methodology approach

Four strains of probiotics were evaluated for their alpha-galactosidase activity. Lactobacillus acidophilus FTCC 0291 displayed the highest specific alpha-galactosidase activity and was thus selected to be optimized in soy whey medium supplemented with seven nitrogen sources. The first-order model showed that meat extract, vegetable extract, and peptone significantly (P < 0.05) influenced the growth of L. acidophilus. The second-order polynomial regression estimated that maximum growth was obtained from the combination of 7.25% (w/v) meat extract, 4.7% (w/v) vegetable extract, and 6.85% (w/v) peptone. The validation experiment showed that response surface methodology was reliable with a variation of only 1.14% from the actual experimental data. Increased utilization of oligosaccharides and reducing sugars contributed to increased growth of L. acidophilus in the soy whey medium. This was accompanied by increased production of short-chain fatty acids and a decrease in pH.     

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。     

Production of a novel 9,12-dihydroxy-10(E)-eicosenoic acid from eicosenoic acid by Pseudomonas aeruginosa PR3

Microbial conversions of unsaturated fatty acids often generate polyhydroxy fatty acids, giving them new properties such as higher viscosity and reactivity. A bacterial strain Pseudomonas aeruginosa (PR3) has been intensively studied to produce mono-, di-, and trihydroxy fatty acids from different 9-cis-monoenoic fatty acids such as oleic acid, ricinoleic acid, and palmitoleic acid. However, from the results and the postulated similar metabolic pathways involved in these transformations, it was assumed that the enzyme system involved in transformation of the monoenoic fatty acid by strain PR3 could utilize fatty acids with different chain lengths and locations of the double bond. In this study was used as a substrate for bioconversion by strain PR3 eicosenoic acid (C20:1, ω-9) containing a singular cis double bond at different positions from the carboxyl end as oleic acid, and it was confirmed that PR3 could produce a novel 9,12-dihydroxy-10(E)-eicosenoic acid (DED) with 6.2% yield from eicosenoic acid. The structure of DED was confirmed using GC-MS, FTIR, and NMR analyses. DED production was maximized at 72 h after the substrate was added to the 24 h culture. Some other nutritional factors were also studied for optimal production of DED.