Evaluation of Cashew Apple Juice for the Production of Fuel Ethanol

A commercial strain of Saccharomyces cerevisiae was used for the production of ethanol by fermentation of cashew apple juice. Growth kinetics and ethanol productivity were calculated for batch fermentation with different initial sugar (glucose + fructose) concentrations. Maximal ethanol, cell, and glycerol concentrations were obtained when 103.1 g L⁻¹ of initial sugar concentration was used. Cell yield (Y _X/S) was calculated as 0.24 (g microorganism)/(g glucose + fructose) using cashew apple juice medium with 41.3 g L⁻¹ of initial sugar concentration. Glucose was exhausted first, followed by fructose. Furthermore, the initial concentration of sugars did not influence ethanol selectivity. These results indicate that cashew apple juice is a suitable substrate for yeast growth and ethanol production.     

Sustainable Surfactin Production by Bacillus subtilis Using Crude Glycerol from Different Wastes

Most biosurfactants are obtained using costly culture media and purification processes, which limits their wider industrial use. Sustainability of their production processes can be achieved, in part, by using cheap substrates found among agricultural and food wastes or byproducts. In the present study, crude glycerol, a raw material obtained from several industrial processes, was evaluated as a potential low-cost carbon source to reduce the costs of surfactin production by Bacillus subtilis #309. The culture medium containing soap-derived waste glycerol led to the best surfactin production, reaching about 2.8 g/L. To the best of our knowledge, this is the first report describing surfactin production by B. subtilis using stearin and soap wastes as carbon sources. A complete chemical characterization of surfactin analogs produced from the different waste glycerol samples was performed by liquid chromatography–mass spectrometry (LC-MS) and Fourier transform infrared spectroscopy (FTIR). Furthermore, the surfactin produced in the study exhibited good stability in a wide range of pH, salinity and temperatures, suggesting its potential for several applications in biotechnology.     

Surfactin production from potato process effluent by Bacillus subtilis in a chemostat

The biosurfactant surfactin has potential to aid in the recovery of energy resources (oil recovery) or subsurface organic contaminants (environmental remediation). However, high medium and purification costs limit its use in these high-volume applications. In previous work, we showed that surfactin could be produced from an inexpensive low-solids potato process effluent with minimal amendments or pretreatments. Previous research has also shown that surfactin can be both produced in Bacillus subtilis cultures and recovered by foam fractionation in an airlift reactor. Results using both purified potato starch and unamended low-solids potato process effluent as substrates for surfactin production indicate that the process is oxygen limited and that recalcitrant indigenous bacteria in the potato process effluent hamper continuous surfactin production. The research reported here features the use of a chemostat operated in batch mode for producing surfactin with concomitant use of antifoam to prevent surfactant loss. The antifoam did not interfere with surfactin recovery by acid precipitation or its efficacy. Initial trials took about 48 h to produce 0.9 g/L of surfactin from potato process effluent. Increasing the oxygen mass transfer by increasing the stirring speed and adding a baffle decreased production time to 12–24 h and produced about 0.6 g/L of surfactin from two different potato-processing facilities.     

Ethanol Production by Fermentation Using Immobilized Cells of Saccharomyces cerevisiae in Cashew Apple Bagasse

In this work, cashew apple bagasse (CAB) was used for Saccharomyces cerevisiae immobilization. The support was prepared through a treatment with a solution of 3% HCl, and delignification with 2% NaOH was also conducted. Optical micrographs showed that high populations of yeast cells adhered to pre-treated CAB surface. Ten consecutive fermentations of cashew apple juice for ethanol production were carried out using immobilized yeasts. High ethanol productivity was observed from the third fermentation assay until the tenth fermentation. Ethanol concentrations (about 19.82–37.83 g L^?1 in average value) and ethanol productivities (about 3.30–6.31 g L^?1 h^?1) were high and stable, and residual sugar concentrations were low in almost all fermentations (around 3.00 g L^?1) with conversions ranging from 44.80% to 96.50%, showing efficiency (85.30–98.52%) and operational stability of the biocatalyst for ethanol fermentation. Results showed that cashew apple bagasse is an efficient support for cell immobilization aiming at ethanol production.     

Ethanol Production from Cashew Apple Bagasse: Improvement of Enzymatic Hydrolysis by Microwave-Assisted Alkali Pretreatment

In this work, the potential of microwave-assisted alkali pretreatment in order to improve the rupture of the recalcitrant structures of the cashew able bagasse (CAB), lignocellulosic by-product in Brazil with no commercial value, is obtained from cashew apple process to juice production, was studied. First, biomass composition of CAB was determined, and the percentage of glucan and lignin was 20.54 ± 0.70% and 33.80 ± 1.30%, respectively. CAB content in terms of cellulose, hemicelluloses, and lignin, 19.21 ± 0.35%, 12.05 ± 0.37%, and 38.11 ± 0.08%, respectively, was also determined. Results showed that, after enzymatic hydrolysis, alkali concentration exerted influence on glucose formation, after pretreatment with 0.2 and 1.0 mo L⁻¹ of NaOH (372 ± 12 and 355 ± 37 mg g_glucan⁻¹) when 2% (w/v) of cashew apple bagasse pretreated by microwave-assisted alkali pretreatment (CAB-M) was used. On the other hand, pretreatment time (15–30 min) and microwave power (600–900 W) exerted no significant effect on hydrolysis. On enzymatic hydrolysis step, improvement on solid percentage (16% w/v) and enzyme load (30 FPU g_CAB-M⁻¹) increased glucose concentration to 15 g L⁻¹. The fermentation of the hydrolyzate by Saccharomyces cerevesiae resulted in ethanol concentration and productivity of 5.6 g L⁻¹ and 1.41 g L⁻¹ h⁻¹, respectively.     

Influence of the Medium Composition and the Culture Conditions on Surfactin Biosynthesis by a Native Bacillus subtilis natto BS19 Strain

An effective microbial synthesis of surfactin depends on the composition of the culture medium, the culture conditions and the genetic potential of the producer strain. The aim of this study was to evaluate the suitability of various medium components for the surfactin producing strain and to determine the impact of the culture conditions on the biosynthesis of surfactin isoforms by the newly isolated native strain Bacillus subtilis natto BS19. The efficiency of surfactin biosynthesis was determined by measuring the surface tension of the medium before and after submerged culture (SmF) and by qualitative and quantitative analysis of the obtained compound by high performance liquid chromatography. The highest efficiency of surfactin biosynthesis was achieved using starch as the carbon source and yeast extract as the nitrogen source at pH 7.0 and 37 °C. Potato peelings were selected as an effective waste substrate. It was shown that the increase in the percentage of peel extract in the culture medium enhanced the biosynthesis of surfactin (mg/L) (2–30.9%; 4–46.0% and 6–58.2%), while reducing surface tension of the medium by about 50%. The obtained results constitute a promising basis for further research on biosynthesis of surfactin using potato peelings as a cheap alternative to synthetic medium components.     

拉曼光谱法快速检测掺入梨汁的浓缩苹果汁

应用便携式拉曼光谱仪结合化学计量学技术建立了浓缩苹果汁掺入梨汁的快速检测方法.实验表明,苹果汁和梨汁的拉曼特征频率相同,给出了特征频率及其对应的分子振动模式.发现苹果汁和梨汁在866 cm-1和1 126 cm-1处的拉曼光谱有微小差别,这可能由于苹果汁和梨汁中果糖异构体含量不同所致.用支持向量分类机(SVC)中的4个核函数对19个苹果汁、梨汁样本进行有效鉴别,准确率达100%,并使用支持向量回归机(SVR)对掺入不同含量梨汁的苹果汁样本进行建模预测,大部分样品相对误差在10%以内.     

Enzymatic Hydrolysis and Fermentation of Pretreated Cashew Apple Bagasse with Alkali and Diluted Sulfuric Acid for Bioethanol Production

The aim of this work was to optimize the enzymatic hydrolysis of the cellulose fraction of cashew apple bagasse (CAB) after diluted acid (CAB-H) and alkali pretreatment (CAB-OH), and to evaluate its fermentation to ethanol using Saccharomyces cerevisiae. Glucose conversion of 82?±?2 mg/g CAB-H and 730?±?20 mg/g CAB-OH was obtained when 2% (w/v) of solid and 30 FPU/g bagasse was used during hydrolysis at 45 °C, 2-fold higher than when using 15 FPU/g bagasse, 44?±?2 mg/g CAB-H, and 450?±?50 mg/g CAB-OH, respectively. Ethanol concentration and productivity, achieved after 6 h of fermentation, were 20.0?±?0.2 g L^?1 and 3.33 g L^?1 h^?1, respectively, when using CAB-OH hydrolyzate (initial glucose concentration of 52.4 g L^?1). For CAB-H hydrolyzate (initial glucose concentration of 17.4 g L^?1), ethanol concentration and productivity were 8.2?±?0.1 g L^?1 and 2.7 g L^?1 h^?1 in 3 h, respectively. Hydrolyzates fermentation resulted in an ethanol yield of 0.38 and 0.47 g/g glucose with pretreated CAB-OH and CAB-H, respectively. Ethanol concentration and productivity, obtained using CAB-OH hydrolyzate, were close to the values obtained in the conventional ethanol fermentation of cashew apple juice or sugar cane juice.     

The Surfactin and Lichenysin Isoforms Produced by Bacillus licheniformis HSN 221

Surfactin and lichenysin are typical members of the lipopeptide family. Four surfactin and two lichenysin isoforms were isolated by Pre RP-HPLC from the cell-free broth of Bacillus licheniformis HSN 221 cultivated in YPD medium. The molecular weight of each isoform was determined by ESI-MS. The peptide part of each isoform was elucidated by Q-Tof MS/MS combined with HPLC, and the fatty acid part was analyzed by GC/MS. It showed that the four surfactin isoforms have the primary structures of anteiso(iso)C₁₃, isoC₁₄, anteiso(iso)C₁₅, anteiso(iso)C₁₅-surfactin-methyl ester, and the two lichenysin isoforms have those of nC₁₄ and anteiso(iso)C₁₅, respectively.     

Production of Microbial Surfactants from Oily Sludge-Contaminated Soil by Bacillus subtilis DSVP23

The indigenous microbial community utilizing aliphatic, aromatic, and polar components from the oily sludge as sole source of carbon and energy was selected from the soil samples of Ankleshwar, India for biosurfactant production. Evaluation of biosurfactant production was done using screening assays such as surface tension reduction, hemolytic activity, emulsification activity, drop-collapse assay, and cell surface hydrophobicity studies. Maximum biosurfactant (6.9?g/l) production was achieved after 5?days of growth from Bacillus subtilis DSVP23 which was identified by 16S RNA technique (NCBI GenBank accession no. EU679368). Composition of biosurfactant showed it to be lipopeptide in nature with 15.2% protein content and 18.0% lipid content. Functional group analysis was also done by using Fourier transform infrared spectroscopy which showed it to be a protein-bound lipid thereby imparting them special properties. Analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometric and nuclear magnetic resonance revealed that the major constituents of lipopeptide are leucine and isoleucine. Gas chromatographic analysis data indicated that oily sludge components of chain length C₁₂?CC₃₀ and aromatic hydrocarbons were degraded effectively by B. subtilis DSVP23 after 5?days of incubation. These results collectively points toward the importance of B. subtilis DSVP23 as a potential candidate for bioremediation studies.     

Synthesis and Evaluation of a Molecularly Imprinted Polymer for Pre-concentration of Patulin from Apple Juice

Patulin, (4-hydroxy-4H-furo[3,2-c] pyran-2(6H)-one) is a toxic secondary metabolite produced by a wide range of fungal species growing on some fruits, including apples. A novel molecularly imprinted polymer (MIP) for patulin has been synthesized using oxindole as a dummy template. The synthesis of MIPs based on dummy templates is a solution to overcome “template bleeding” shortcoming in trace analysis. The polymer was prepared in a non-covalent approach with methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker via free radical polymerization. It was revealed that the MIP particles in extraction columns displayed high affinity towards patulin and, therefore, subsequently employed for molecularly imprinted-solid phase extraction (MIP-SPE) of patulin from contaminated apple juice samples. The analysis of spiked samples showed good recoveries (>80%). Reproducibility, repeatability, and limit of detection of the proposed method were also investigated.     

高效液相色谱测定浓缩苹果汁展青霉素的残留量

报道采用高效液相色谱(HPLC)法分析浓缩苹果汁中展青霉素的含量。果汁样品先经纯水稀释,再用乙酸乙酯提取,通过净化、浓缩,用甲醇/水(1∶1,V/V)定容后,进行HPLC检测。色谱柱为资生堂MG-ⅡC18柱,采用等度洗脱,流动相为甲醇、水、乙酸-乙酸钠盐缓冲液(pH=4.6);采用可变波长检测器(VWD)在276 nm波长检测。展青霉素在0.02~1.0μg/mL范围内与其峰面积呈良好的线性关系,其相关系数(R2)为0.9965。果汁样品中不同水平标准加入回收率在69.82±1.64%~87.54±3.61%范围;展青霉素在样品中残留量检测的定量限为50μg/L。     

Optimization of Surfactin Production by <Emphasis Type="Italic">Bacillus subtilis</Emphasis> Isolate BS5

Bacillus subtilis BS5 is a soil isolate that produces promising yield of surfactin biosurfactant in mineral salts medium (MSM). It was found that cellular growth and surfactin production in MSM were greatly affected by the environmental fermentation conditions and the medium components (carbon and nitrogen sources and minerals). Optimum environmental conditions for high surfactin production on the shake flask level were found to be a slightly acidic initial pH (6.5-6.8), an incubation temperature of 30 degrees C, a 90% volumetric aeration percentage, and an inoculum size of 2% v/v. For media components, it was found that the optimum carbon source was molasses (160 ml/l), whereas the optimum nitrogen source was NaNO(3) (5 g/l) and the optimum trace elements were ZnSO(4).7H(2)O (0.16 g/l), FeCl(3).6H(2)O (0.27 g/l), and MnSO(4).H(2)O (0.017 g/l). A modified MSM (molasses MSM), combining the optimum medium components, was formulated and resulted in threefold increase in surfactin productivity that reached 1.12 g/l. No plasmid could be detected in the tested isolate, revealing that biosurfactant production by B. subtilis isolate BS5 is chromosomally mediated but not plasmid-mediated.     

Self-Cloning Significantly Enhances the Production of Catalase in Bacillus subtilis WSHDZ-01

The katA gene that encodes catalase (CAT) in Bacillus subtilis WSHDZ-01 was overexpressed in B. subtilis WB600 and B. subtilis WSHDZ-01. The CAT yield in both transformed strains was significantly improved compared to that in the wild-type WSHDZ-01 in shake flask culture. When cultured in a 3-L stirred tank reactor (STR), the recombinant CAT activity in B. subtilis WSHDZ-01 could be improved by 419 %, reaching up to 39,117 U/mL and was 8,149.4 U/mg dry cell weight, which is the highest activity reported in Bacillus sp. However, the recombinant CAT in B. subtilis WB600 cultured in a 3-L STR was not significantly improved by any of the common means for process optimization, and the highest CAT activity was 3,673.5 U/mg dry cell weight. The results suggest that self-cloning of the complete expression cassette in the original strain is a reasonable strategy to improve the yield of wild-type enzymes.     

Production of XynX, a Large Multimodular Protein of Clostridium thermocellum, by Protease-Deficient Bacillus subtilis Strains

XynX of Clostridium thermocellum is a large, multimodular xylanase of 116?kDa. An Escherichia coli transformant carrying the entire xynX produced three active truncated xylanase species of 105, 85, and 64?kDa intracellularly. The Bacillus subtilis WB700 transformant with the xynX, a strain deficient in seven proteases including Vpr, secreted two active truncated xylanase species of 65 and 44?kDa. The B. subtilis WB800 transformant with xynX, a strain deficient in eight proteases including Vpr and WprA, secreted more active enzymes, 8.46?U?ml^?1, mostly in the form of 105 and 85?kDa, than the WB700 transformant, 6.93?U?ml^?1. This indicates that the additional deletion of wprA enabled the WB800 to secrete XynX in its intact form. B. subtilis WB800 produced more total enzyme activity than E. coli (1,692?±?274 U vs. 141.9?±?27.1 U), and, more importantly, secreted almost all the enzyme activity. The results suggest the potential use of B. subtilis WB800 as a host system for the production of large multimodular proteins.     

Biosurfactant production by Bacillus subtilis using cassava-processing effluent

A cassava flour-processing effluent (manipueira) was evaluated as a substrate for surfactant production by two Bacillus subtilis strains. B. subtilis ATCC 21332 reduced the surface tension of the medium to 25.9 mN/m, producing a crude biosurfactant concentration of 2.2 g/L. The wild-type strain, B. subtilis LB5a, reduced the surface tension of the medium to 26.6 mN/m, giving a crude biosurfactant concentration of 3.0 g/L. A decrease in surfactant concentration observed for B. subtilis ATCC 21332 seemed to be related to an increase in protease activity. The biosurfactant produced on cassava effluent medium by B. subtilis LB5a was similar to surfactin.     

气相色谱-质谱联用对浓缩苹果汁中多种拟除虫菊酯类农药的快速测定

苹果汁饮料是世界果汁消费市场最主要的产品之一,我国是浓缩苹果汁第一大出口国.由于近年来拟除虫菊酯残留问题日益突出,发达国家对浓缩苹果汁中拟除虫菊酯类农药进行了严格的限量规定.     

断续流动-氢化物发生原子荧光法测定浓缩苹果汁中的砷

用断续流动－氢化物发生原子荧光法测定了浓缩萍果汁中的砷含量。确定了仪器的最佳条件，探讨了酸介质，硫脲－抗坏血酸用量，硼氢化钾加入量对测定As的影响。在选定的操作条件下，砷的检限为0．43ng/mL，相对标准偏差为3．4％－6．3％，加标回收率为94．0％－101．5％本方法简便，快速，灵敏，准确，应用于浓缩苹果汁中As的测定，得到满意的结果。