学生食堂餐厨垃圾中淀粉的生物利用菌株筛选

目的探索地衣芽胞杆菌、枯草芽胞杆菌和蜡样芽胞杆菌分解大学生食堂厨余中淀粉的能力,以筛选和研制餐厨垃圾生物降解的使用菌种。方法将各菌种接于淀粉酶试验培养基,培养后滴加碘溶液,观察透明圈,判定产淀粉酶能力;收集大学生食堂的厨余,观察三种细菌在不同接种量（5%、10%、15%、20%、25%）、不同接种时间（24 h、48 h、72 h）及不同菌株配伍方式下发酵淀粉的能力。结果地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌都能产生淀粉酶,以地衣芽胞杆菌产生的淀粉酶较多,其次为枯草芽胞杆菌,蜡样芽胞杆菌较少,三种细菌分解厨余中淀粉的最佳接种量都为15%-20%,最佳发酵时间为48 h,枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵效果最佳。结论可采用枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵学生食堂的厨余中的淀粉。     

巴氏杀菌对餐厨垃圾高温厌氧发酵的影响

通过对杀菌、未杀菌餐厨垃圾高温发酵对比试验的研究分析,阐述了餐厨垃圾杀菌处理对发酵的影响。杀菌方式为巴氏消毒法,温度70℃,持续时间10 min,接种物为德青源实验室自行培养的高温发酵菌,底物为餐厨垃圾,有机负荷5 gVS/L,污泥负荷(F/M)为0.5,试验持续32 d,结果显示:杀菌组累计产气量895 mL/gVS,未杀菌组累计产气量795 mL/gVS;杀菌样品产气速率高于未杀菌样品,其VS去除率略低于未杀菌样品。     

餐厨垃圾高温菌种对斑马鱼急性毒性与安全评价

目的:研究餐厨垃圾高温菌种对斑马鱼的毒性,为其安全使用提供依据。方法:以具有代表性的水生动物斑马鱼为试材,采用静态法测定餐厨垃圾高温菌种对斑马鱼急性毒性并对其进行安全评价。结果:鱼的半数致死浓度LC50值24、48和96h分别为3400mg/L、3000mg/L和2500mg/L。结论:餐厨垃圾高温菌种对斑马鱼有一定毒性,该研究为餐厨垃圾高温菌种的安全合理使用提供了科学依据。     

外接堆肥微生物在餐厨废弃物好氧堆肥中的应用

微生物是堆肥过程的主要驱动力,其群落在堆肥过程中不断演替,进而影响好氧发酵进程和堆肥的品质。大量研究表明：接种外源微生物可促进堆肥进程,加快有机物分解和堆肥腐熟,消解抗生素等有害物质等作用。餐厨废弃物油脂、盐、水含量高和易酸化等问题会影响好氧发酵过程中微生物的活动。通过特定样品的富集驯化、选择性培养,并结合先进的分子生物学技术,筛选具有不同降解功能的菌株,并以此为基础构建的微生物菌剂能够一定程度上克服餐厨废弃物用于好氧堆肥的限制性问题。主要阐述了餐厨废弃物好氧发酵过程中微生物菌群的演替规律,构建微生物菌剂的菌种类型及功能和不同菌剂对好氧发酵的作用及潜在影响机制,以期能够为相关微生物强化技术的研发提供一些参考。     

西安市餐厨垃圾规范化管理和对策

本文就加强西安市餐厨垃圾管理提出了一些建议和对策,希望对实现西安市餐厨垃圾规范化管理和发展循环经济起到积极作用。     

产油微生物利用餐厨垃圾生产油脂的研究进展

餐厨垃圾中含有丰富的营养物质,经生物转化过程可以合成对人类有用的化学品.某些产油微生物可以处理餐厨垃圾生产油脂,同时合成高附加值代谢产物如多不饱和脂肪酸、角鲨烯和类胡萝卜素等.这不仅能够降低生产成本,而且提高了产物的经济价值,具有极大的工业化应用潜力.文中主要概括了目前餐厨垃圾的处理研究现状,综述了产油微生物发酵餐厨垃...     

餐厨垃圾厌氧消化产沼气过程中酶学表征

厌氧消化产沼气被认为是餐厨垃圾资源化利用的有效方式之一,其实质是在多种微生物综合作用下的生物化学过程.本文研究了在促进和抑制性因子作用下,餐厨垃圾厌氧发酵的酶学过程,对其中的脱氢酶和水解酶(β-葡萄糖苷酶,BAA-蛋白水解酶,碱性磷酸酶)活性变化进行了分析.研究表明,与空白对照组相比添加酵母粉后脱氢酶的最高活性提高了8...     

基于酶促反应的餐厨垃圾厌氧发酵效率优化研究

酶促反应预处理餐厨垃圾可提高甲烷产量,减少有害物质积累,优化发酵体系特性。在协同发酵中,酶促反应可通过调节底物碳氮比值、降低有害物质抑制作用、提高甲烷产量、改善发酵体系物理特性,有效提升餐厨垃圾厌氧发酵的效率。工艺参数优化、微生物群落调控以及酶促反应与两相厌氧发酵的结合,是提高餐厨垃圾厌氧发酵效率的关键技术路径。通过优化这些条件,可提升甲烷产量和有机物转化率,有望为餐厨垃圾资源化利用提供新的技术支持。     

餐厨垃圾资源化利用技术研究现状及展望

餐厨垃圾产生量大、成分复杂、具有“危害性”和“资源性”的双重属性,随着垃圾分类工作的推进,将餐厨垃圾作为一种生物资源回收其中的资源和能源的研究受到了越来越多的关注。本文介绍了餐厨垃圾的成分特性及预分选方法,对餐厨垃圾厌氧消化、好氧堆肥、生物饲料、昆虫养殖、热处理技术及生物炼制生产高附加值化学品等主要的资源化利用途径进行了分析,并对餐厨垃圾收集及资源化过程中产生的恶臭气体、废水污染问题及处理方法进行了介绍。最后指出餐厨垃圾厌氧消化、昆虫养殖、好氧堆肥及生物饲料技术工业化利用过程中的设备运行的稳定性及其废水和臭气的控制问题仍需进一步的研究。餐厨垃圾热处理过程如何进一步降低能耗及开发高附加值的功能炭材料是未来的重要发展方向,餐厨垃圾生物炼制生产高附加值化学品是实现餐厨垃圾高值化利用的有效途径,也是替代传统化工路线生产化学品的重要路径。总之,采用多技术耦合是实现餐厨垃圾“减量化、无害化、资源化”的有效手段,也是发展我国循环经济发展的必然要求。     

湿垃圾高温好氧堆肥处置过程中嗜热菌的应用及研究进展

目前,湿垃圾的无害化处理越来越受到重视,我国湿垃圾的主要处置方式从焚烧和填埋逐渐发展到微生物发酵,其中,好氧堆肥能够有效处理农业废弃物、厨余垃圾以及厌氧发酵存留的沼渣,是多数湿垃圾处置的最终归宿。近年来,随着小型堆肥设施的发展和普及,高温好氧堆肥逐渐成为主流的堆肥方式。相较于传统自然堆肥,高温好氧堆肥技术通过额外添加高温菌剂及其他辅助手段,显著提升了堆肥的温度及其持续时间,有效提高了堆肥质量。优良的高温降解菌的筛选和菌剂配制是高温好氧发酵技术发展的前提。目前,在湿垃圾高温好氧堆肥中,常用的菌种包含各种放线菌、芽孢杆菌、假单胞菌和乳酸菌等。使用不同菌种复配的高温菌剂能够明显延长高温持续时间,提升堆肥中不同组分的降解效率,有效减灭堆肥样本的有害性,使之成为能够二次利用的优良有机腐殖质。本文阐述了目前我国湿垃圾的概况、湿垃圾现有的处置方式、嗜热菌的种类及筛选、高温好氧堆肥的技术特点以及发展趋势等,初步讨论了目前湿垃圾转化面临的机遇与挑战,为湿垃圾的无害化、资源化处理提供一些有价值的参考意见。     

alkB homologs in thermophilic bacteria of the genus Geobacillus

Screening for alkane hydroxylase genes (alkB) was performed in thermophilic aerobic bacteria of the genus Geobacillus. Total DNAs were isolated from the biomass of 11 strains grown on a mixture of saturated C₁₀–C₂₀ hydrocarbons. Fragments of alkB genes were amplified by PCR with degenerate oligonucleotide primers, and the PCR products were cloned and sequenced. For the first time, a set of alkB gene homologs was detected in the genomes of thermophilic bacteria. The strains each contained three to six homologs, of which only two were common for all of the strains. Phylogenetic analysis of the nucleotide sequences and the deduced amino acid sequences showed that six of the variants revealed in Geobacillus were closely related to alkB4, alkB3, and alkB2, found in Rhodococcus erythropolis strains NRRL B-16531 and Q15. All variants of alkB sequences were unique. Analysis of the GC composition showed that the Geobacillus alkB homologs are closer to Rhodococcus than to Geobacillus chromosomal DNA. It was assumed that the alkB genes were introduced in the Geobacillus genome via interspecific horizontal transfer and that Rhodococcus or other representatives of Actinobacteria served as donors. Analysis of the codon usage in the fragments of alkB genes confirmed the suggestion that the pool of these genes is common to the majority of Gram-positive and certain Gram-negative bacteria. The formation of a set of several alkB homologs in a genome of a particular microorganism may result from free gene exchange within this pool.     

Production of a thermophilic,extracellular alkaline protease by Bacillus stearothermophilus AP-4

A thermophilic Bacillus stearothermophilus strain AP-4 excreting a thermostable alkaline protease, was isolated from a local compost. Maximum activity of protease (250 U/ml) was after 36 h growth in broth at pH 9.0 and at 55°C. The protease was optimally active at pH 9.0 and 55°C and was stable in 5 mm CaCl₂. The enzyme was completely inactivated by PMSF, EDTA and -mercaptoethanol. It is therefore a metal ion-dependent, alkaline, serine protease.R. Dhandapani and R. Vijayaragavan are with the Centre for Plant Molecular Biology & Biotechnology, Tamil Nadu Agricultural University, Coimbatore 641 003, India     

Biotransformation of halophenols by a thermophilic Bacillus sp.

The thermophilic Bacillus sp. A2 transformed various halophenols. 2-Chlorophenol, 2-bromophenol, 3-bromophenol and 2-fluorophenol were transformed under resting cell conditions at 60°C to 3-chlorocatechol, 3-bromocatechol, 4-bromocatechol and 3-fluorocatechol, respectively. The hydroxylation of 3-bromophenol occurred at the proximal and distal position relative to the halogen substituent. In complex medium this strain completely transformed 2-chlorophenol and 2-bromophenol at concentrations up to 1 mM. Concomitantly, an accumulation of oxygen-and temperature sensitive halocatechols was observed. 3-Chlorocatechol possesses a half-life of 11.5 h at 60°C and is therefore readily decomposed during incubation. The hydroxylating system was present in phenolgrown cells but not in glucose-grown cells. The hydroxylase activity could also be induced by 2-chlorophenol. The product, 3-chlorocatechol, is not a substrate for the catechol 2,3-dioxygenase.Abbreviations 2-CP 2-chlorophenol - DCP dichlorophenol - TCP trichlorophenol - tetraCP tetrachlorophenol - MIC minimal inhibitory concentration - CF chloride-free - CFG chloride-free plus glucose - CFGY chloride-free plus glycerol - CFP chloride-free plus phenol - CAM chloramphenicol     

一株深海热液口嗜热芽孢杆菌SY27F代谢产物活性的研究

【目的】对一株来源于深海热液口嗜热芽孢杆菌的次生代谢产物进行抑菌活性和抗肿瘤活性的初步研究。【方法】采用纸片法和微量肉汤稀释法检测嗜热芽孢杆菌SY27F次生代谢产物的抑菌活性,采用CCK-8法测定其次生代谢产物的抗肿瘤活性。【结果】抑菌实验表明,嗜热芽孢杆菌代谢产物对大肠杆菌、金黄色葡萄球菌均有抑菌作用,其最低抑菌浓度分别为1.56 mg/mL和3.13 mg/mL;细胞实验表明,其代谢产物对肿瘤细胞A549、HepG2、HeLa、MCF-7均有一定的抑制作用,其半致死浓度分别为0.390、0.451、0.704、1.105 mg/mL;与人肝肿瘤细胞(HepG2)相比,其对人正常肝细胞(L02)表现出良好的生物相容性。【结论】嗜热芽孢杆菌SY27F次生代谢产物具有一定的抑菌和抗肿瘤活性,可为寻找新型抑菌抗肿瘤活性物质提供优质资源。     

A Catecholic Siderophore Produced by the Thermoresistant Bacillus licheniformis VK21 Strain

Thermophilic and thermoresistant strains of bacilli were screened on a medium containing Chrome Azurol S for the producers of siderophores. It was found that the Bacillus licheniformis VK21 strain dramatically increases secretion of the metabolite, a chelator of Fe³⁺, in response to addition of manganese(II) salts. The growth of the producer on a minimal medium containing MnSO₄ under the conditions of iron deficiency is accompanied by the accumulation of a catecholic product, the content of which reaches maximum at the beginning of the stationary growth phase of culture. In the presence of FeCl₃, the amount of the catecholic product in the medium considerably decreases. The siderophore, called S_VK21, was isolated from the cultural medium and purified by reversed phase HPLC, and its siderophore function was confirmed by the test for the restoration of growth of producer cells in a medium containing EDTA. The UV spectrum of the siderophore has absorption maxima at 248 and 315 nm. According to the amino acid analysis and NMR spectrometry, the metabolite S_VK21 is 2,3-dihydroxybenzoyl-glycyl-threonine.     

Expression of the cloned xylanases from an alkalophilic,thermophilic Bacillus in Bacillus subtilis

A recombinant plasmid construct, pLPX6.5, harbouring a 6.5 kb Hind III fragment of genomic DNA, from an alkalophilic, thermophilic Bacillus NCIM 59 and coding for xylanase activity, was electroporatically transformed into Bacillus subtilis MI 111. The expression of the recombinant xylanases was confirmed by cross-reactivity with antibodies raised against purified xylanase II (M _r 15,800) from NCIM 59. However, as there were different xylan hydrolysis products from NCIM 59 and the host B. subtilis, the two xylanases appear to have different modes of action. Xylanase expression in the transformants was 6-fold higher than in the host. There was no significant enhancement in the expression of recombinant xylanases by adding xylan to the growth medium.The authors are with the Division of Biochemical Sciences, National Chemical Laboratory, Pune-411008, India     

Intensive bioconversion of sewage sludge and food waste by Bacillus thermoamylovorans

The main aim of this work was to intensify conventional composting of a mixture of sewage sludge and solid food wastes by a one-stage thermophilic bioconversion of these wastes into an organic fertilizer. An intensive process was carried out in a closed system, with or without addition of a starter culture of Bacillus thermoamylovorans. The most effective thermophilic bioconversion of the mixture of food waste and sewage sludge, with addition of starter culture, was when the pH was buffered with calcium carbonate, or the pH drop in the material was prevented by preliminary removal of sulphides from sewage sludge by hydrogen peroxide.     

Constitutive production of endoglucanase by a Bacillus sp. isolated from a Zimbabwean hot spring

A sporulating, aerobic Bacillus sp., isolated from Chimanimani hot springs, Zimbabwe, produced endoglucanase when cultured on medium with initial pH between 5.0 and 9.0 and at 30 to 60°C. Optimal production of endoglucanase was at pH 6.0. The enzyme was constitutively produced when the organism was cultured on starch, cellobiose, carboxymethylcellulose, sucrose, glucose, galactose, Avicel, lactose, mannose or maltose.The authors are with the Fermentation and Food Group, Department of Biochemistry, University of Zimbabwe, Box MP 167, Mount Pleasant, Harare, Zimbabwe     

Two acidothermotolerant lipases from new variants of Bacillus spp.

Two acidothermotolerant lipases from new isolates of Bacillus stearothermophilus SB-1 and Bacillus licheniformis SB-3 are reported. In addition, a thermotolerant, neutral lipase from Bacillus atrophaeus SB-2 that hydrolyses castor oil is also reported. The lipase from B. stearothermophilus SB-1 retained 70% activity and that from B. licheniformis SB-3 retained 50% activity at pH 3.0 at 50 °C. In addition, at 100 °C B. stearothermophilus SB-1 lipase had a half life of 25 min at pH 3.0 and 15 min at pH 6.0. Lipase activity was markedly stimulated by glycerol in case of B. stearothermophilus SB-1 and by diethylether in cases of B. atrophaeus SB-2 and B. licheniformis SB-3. The lipases varied in their substrate specificity towards triacylglycerols. The rate of hydrolysis of neem oil with B. stearothermophilus SB-1 and B. atrophaeus SB-2 lipases was, respectively, nearly 4-fold and 2-fold more than with olive oil.     

Conversion of food waste into hydrogen by thermophilic acidogenesis

Conversion of food waste into hydrogen by thermophilic acidogenesis was investigated as a function of organic loading rate (OLR), hydraulic retention time (HRT) and pH in a continuous stirred tank reactor. In order to identify hydrogen-producing microorganisms, denaturing gradient gel electrophoresis (DGGE) of the polymerase chain reaction (PCR) – amplified V3 region of 16S rDNA analysis was conducted at each tested pH. The conversion of food waste into hydrogen was strongly influenced by the operational conditions. The hydrogen production was increased as OLR increased up to 8gVSl^-1d^-1, but drastically decreased at 10gVSl^-1d^-1. The yield of hydrogen was decreased from 2.2 to 1.0mol-H₂/mol-hexose consumed as HRT decreased from 5 to 2days. More carbohydrates in the food waste were decomposed at longer HRT, 76–90%, at HRT of 2–5days. The hydrogen production peaked at pH 5.5±0.1 and significantly decreased at pH 5.0±0.1. The biogas produced was composed of hydrogen and carbon dioxide, but no methane was detected at all tested conditions. The hydrogen contents in the gas produced were more than 55% (v/v) and not sensitive to all tested conditions. The optimum operational condition for continuous hydrogen production from the food waste was obtained at 8gVSl^-1d^-1, 5 days HRT and pH 5.5±0.1 where the hydrogen production rate, content, yield and the efficiency of carbohydrate decomposition were 1.0l H₂/l-d, 60.5% (v/v), 2.2mol-H₂/mol-hexose consumed and 90%, respectively. The hydrogen production was related with the concentration of total organic acids (TOA) which was strongly dependent on that of butyrate indicating that the reaction was mainly butyrate fermentation. The hydrogen-producing microorganism of Thermoanaerobacterium thermosaccharolyticum that involved in acetate/butyrate fermentation, was detected with strong intensity at all tested pHs by denaturing gradient gel electrophoresis (DGGE) of the polymerase chain reaction (PCR) – amplified V3 region of 16S rDNA analysis and sensitive to the tested pHs. The experimental results indicated that effective hydrogen production from the food waste could be obtained continuously by thermophilic acidogenesis at proper operational condition.