混合培养微生物利用甘油补料发酵生产乙醇研究

采用浸麻芽孢杆菌和红曲菌990691用甘油混合发酵生产乙醇.结果表明,分批发酵中高浓度的甘油对乙醇发酵有着较强的抑制作用,分批发酵最佳甘油浓度为0.217 mol L-1.在分批发酵的基础上补料发酵,考察了不同甘油浓度的补料液和不同补料时间对乙醇发酵的影响.最终确定乙醇补料发酵较优的工艺条件为:反应器1 L,装液量700 mL红曲发酵液,甘油初始浓度为0.217 mol L-1,以补料方式每隔60 h分5次补加0.217 mol L-1甘油浓度的红曲发酵液,每次补加100 mL,发酵培养360 h.当乙醇最高浓度达0.221 mol L-1,乙醇总产率0.628 mmol h-1,乙醇/甘油转化率达87%(mol mol-1).与分批发酵相比,补料发酵很大程度解除了高浓度甘油的抑制作用,有效地利用了甘油,提高了乙醇的产量,且乙醇产率较为稳定.     

Optimization of fermentation process for saccharide-based fuel ethanol production by Saccharomyces cerevisiae北大核心CSCD

低成本、高产量的发酵工艺是实现工业燃料乙醇经济和环境可持续性发展的关键,而不需要重大基础设施改变或投资.为获得酿酒酵母(Saccharomyces cerevisiae)利用甘蔗汁生产燃料乙醇的最优发酵工艺,首先对发酵体系的氮源条件进行优化;其次,在单因素试验基础上,以乙醇发酵效率为响应值,通过响应面法优化了燃料乙醇生产的发酵工艺,并通过补料分批发酵技术在5 L发酵罐中进一步扩大发酵.结果表明,以1.0 g/L (NH)SO和1.0 g/L酵母提取物作为发酵氮源,乙醇发酵效率和得率比对照可分别提高4.80%、9.52%.响应面设计获得的最优发酵工艺条件为在总糖浓度150.0 g/L、酵母提取物浓度2.0 g/L、发酵时间24.5 h、pH5.0、外加(NH)SO浓度1.0 g/L时,最高乙醇发酵效率可达到91.10%.在5 L发酵罐中采用补料分批发酵获得的最终乙醇浓度达到98.92 g/L,发酵效率维持在90%左右,乙醇生产力最高达到3.81 g Lh.本研究获得了一种高效生产糖质燃料乙醇的发酵工艺,可在较短时间内获得高浓度乙醇且消耗较少氮源,结果可为进一步利用糖质原料进行高效生物炼制及高浓度乙醇工业化生产提供参考.(图6表6参30)     

低pH值下上流式厌氧污泥床反应器(UASB)以糖蜜为底物制取高纯度氢气

采用由有机玻璃制成的上流式厌氧污泥床反应器(UASB),以糖蜜废水为发酵底物,投加适量氯化铵和磷酸二氢钾,研究在低p H值下系统的产氢性能.在系统启动初期,投加碳酸氢钠控制进水p H=6.75—7.15,使系统处于中性厌氧发酵,固定进水COD为4000 mg·L~(-1),HRT=8 h.系统运行20 d后,系统处于混合型发酵类型,随后停止投加碳酸氢钠.系统经过45 d的运行,乙醇和乙酸浓度占总浓度的79.39%,而丙酸的浓度只占总浓度的10.89%,形成了稳定的典型乙醇型发酵.第65天,乙醇和乙酸的浓度分别为840.56 mg·L~(-1)、403.12 mg·L~(-1),乙醇与乙酸浓度占总浓度的93.2%,氢气含量为86.97%.在出水p H=2.81时,系统产氢性能最佳.氢气产率为2.079 mmol·L~(-1)·h~(-1),氢气产量1.12 m3·m-3·d~(-1),氢气含量91.46%,是稳定期氢气含量的1.65倍.     

初期通气和震荡培养提高高浓度乙醇发酵的乙醇浓度和产率

研究了氧气和震荡条件对酿酒酵母高浓度乙醇发酵的影响.结果表明,震荡是提高发酵液乙醇浓度和产率的最重要因素.与静止培养相比,在不通气情况下震荡培养使乙醇浓度提高了69%(从75.8 g L-1提高到128.1 g L-1),在通气条件下乙醇浓度提高了68.7%(从85.2 g L-1提高到to 143.8 g L-1).在最优条件下,两次补料,经54 h发酵,发酵液中乙醇浓度达到143.8 g L-1,乙醇产率与理论产率的比值为0.471 g/g(即92.20%).经分析,通气和震荡条件提高了发酵液中酿酒酵母的生物量和细胞活力.图5表1参12     

鲜甘薯发酵生产高浓度乙醇的技术

乙醇作为燃料可以改善能源结构,减少对石油进口的依赖.高浓度发酵技术是一种生产燃料乙醇的新兴技术,有利于降低乙醇的生产成本.本研究采用酿酒酵母以鲜甘薯为底物进行了快速高浓度乙醇发酵的研究.对高浓度乙醇发酵的影响因素如发酵促进剂种类和浓度、无机盐、维生素及初糖浓度进行了探讨,获得了最佳发酵培养基配方,确定最适发酵促进剂为B,浓度为1.20 g kg-1,不需要添加无机盐和维生素,初糖浓度为270 g kg-1.在最适条件下,28 h可生产乙醇132.86 g kg-1,乙醇发酵强度达4.74 g kg-1h-1,发酵效率达91.44%.发酵规模放大至10 L时,28 h可产生乙醇131.71 g kg-1,乙醇发酵强度为4.70 g kg-1h-1,发酵效率为90.53%.图3表3参16     

菊芋秸秆高浓度物料分步糖化及乙醇发酵

菊芋是生物能源和生物炼制的新型原料作物,具有和其他作物不同的秸秆组成.为了解菊芋秸秆的生物转化情况,本研究首先比较了NaOH-H_2O_2、瞬间弹射蒸汽爆破(ICSE)及NaOH-H_2O_2和ICSE联用等3种预处理方法,证明对于菊芋秸秆NaOH-H_2O_2预处理法简单高效.进一步研究显示,NaOH-H_2O_2预处理过程中水洗一次即可显著促进酶解和后续发酵.利用分批补料和补加纤维素酶的方式进行高物料浓度条件下预处理菊芋秸秆的分步水解和乙醇发酵,当物料浓度达到30%(m/V)时,水解72 h的葡萄糖和木糖浓度分别可达143.6 g/L和36.2 g/L.利用木糖-葡萄糖共发酵重组酿酒酵母菌株LX03在菊芋秸秆水解液中进行乙醇发酵,发酵72 h乙醇最高浓度达66.2 g/L(8.27%,V/V),且发酵总糖利用率达86.9%.本研究利用菊芋秸秆水解液发酵获得较高的乙醇产量,为进一步利用菊芋秸秆进行高效生物炼制及高浓度纤维素乙醇生产提供了参考.(图3表1参23)     

一株运动发酵单胞菌Zy-1快速生产乙醇

经多次实验优化，得到运动发酵单胞菌Zy发酵葡萄糖生产乙醇较合适的条件．Zy的诱变菌株Zy-1在该条件下发酵葡萄糖生产乙醇比原始菌株更有较大优势．当葡萄糖浓度为200gL^-1时，发酵48h，乙醇浓度96．5gL^-1，残糖2．3gL^-1，发酵效率为94．42％．Zy-1发酵天然原料米粉、木薯、红薯干等，发酵时间44h，乙醇浓度达95gL^-1以上，发酵效率92％以上．发酵液用DNS法测定，还原糖约2gL^-1，残总糖因原料种类不同，其值有所差异（5～20gL^-1）．经薄层层析分析，发酵液无葡萄糖，而是二糖、三糖等低聚糖．图2表5参12     

鲜甘薯原料的运动发酵单胞菌快速乙醇发酵条件

对运动发酵单胞菌232B同步糖化发酵(SSF)鲜甘薯快速生产燃料乙醇的条件进行了研究.通过单因素试验和正交试验获得了乙醇发酵的最佳参数为:初始pH值6.0～7.0,硫酸铵5.0 g/kg,糖化酶量1.6 AUG/kg淀粉,初始总糖浓度200 g/kg,接种量ψ=12.5%.经过21 h发酵,乙醇浓度为95.15 g/kg.发酵效率可达94%.同时对不灭菌发酵也进行了研究,发酵效率可达92%.残糖的HPLC分析结果说明,发酵液中已没有葡萄糖存在,经酸水解后又出现了葡萄糖、半乳糖、甘露糖等成分,说明发酵结束后的残糖是多种低聚糖.图4表4参19     

混合糖发酵条件下甲酸抑制木糖发酵的机制

纤维素燃料乙醇生产面临的一个重要问题是纤维素原料预处理过程中产生多种副产物会显著抑制酿酒酵母的生长繁殖和发酵,其主要成分弱酸类中甲酸被认为具有最强的抑制效应.为了解工业酿酒酵母混合糖发酵时木糖利用被甲酸特异性显著抑制的机制,为发酵菌株的抑制物耐受育种提供依据,以乙酸存在条件下的发酵为对照,研究甲酸存在条件下菌株分别发酵混合糖和单独木糖时的发酵性能以及糖代谢相关基因的表达差异、葡萄糖浓度对菌株发酵木糖和木糖代谢基因表达的影响,同时研究甲酸存在条件下葡萄糖代谢产物乙酸和乙醇对菌株发酵木糖的影响以及混合糖和单独木糖发酵过程中甲酸浓度变化和甲酸脱氢酶基因FDH1转录情况.结果显示:葡萄糖只有在甲酸存在条件下才特异性地显著抑制木糖发酵,木糖消耗速率的下降与木糖还原酶(XR)和木糖醇脱氢酶(XDH)酶活下降有关;单独木糖发酵时,只有当乙酸、乙醇和甲酸共存时才表现出抑制效应,且随乙醇浓度增加抑制效应越明显,木糖发酵被抑制与XDH酶活下降有关,但乙酸、乙醇和甲酸三者对木糖发酵的协同抑制效应明显弱于60 g/L葡萄糖存在时的抑制;混合糖发酵时FDH1基因转录被抑制导致甲酸分解缓慢,对甲酸存在条件下木糖发酵被抑制有部分贡献.综上,葡萄糖抑制甲酸分解与葡萄糖代谢产物乙酸、乙醇和甲酸的协同抑制对混合糖发酵时甲酸对木糖发酵特异性显著抑制有贡献,但尚存在其他未知抑制机制,还需进一步深入研究.     

混合菌群产氢特性研究

对产氢菌株进行筛选,得到一组可以在微氧条件下高效产氢的微生物菌群.此菌群在0～15%O2浓度下都可以产氢,具有较高的耐氧产氢特性.该混合菌群可利用甘露醇、葡萄糖、蔗糖、乳糖、淀粉为底物产氢,其中甘露醇为最适底物.最适产氢温度、pH值、仞始氧气浓度分别为33℃、7.0、2.72%.在此条件下,以片露醇为碳源(5.0 g/L),产氢效率可达到324.18 mL(H2)/g(甘露醇).对该产氢体系发酵末端产物的液相分析显示乙醇占76%～93%,表明该产氢体系为乙醇型发酵.通过PCR-DGGE方法进行菌群分析,发现不同初始氧浓度下菌群分布有一定差异,但克氏杆菌在各种氧浓度下的混合菌群中都占明显优势,是主要的产氢菌.图8表1参25     

Rapid,recurring, structured survey versus bioblitz for generating biodiversity data and analysis with a multispecies abundance model

A bioblitz inexpensively and quickly generates biodiversity data, but bioblitzes are often conducted with haphazard, unreplicated sampling. Results tend to be taxonomically, geographically, or temporally biased, lack metadata, and consist of lists of observed taxa that do not enable further analyses or correction for imperfect detection. A rapid, recurring, structured survey (RRSS) uses a structured sampling design and temporal and spatial replication to survey randomly selected sites on a conservation property. We participated in a loosely structured bioblitz and a subsequent RRSS at Big Canoe Creek Nature Preserve in Springville (St. Clair County), Alabama (USA) to compare observed richness derived from the 2 survey approaches. The RRSS data structure enabled us to fit models that accounted for imperfect detection to estimate abundances, occupancy probabilities, and habitat associations. The loosely structured bioblitz data could not be used in such models. We present a new integrated multispecies abundance model that we applied to avian RRSS data. Our model extension enables estimation for the community, employs data augmentation to estimate the number of undetected species, and incorporates covariates. The RRSS generated a more comprehensive and less biased list of observed taxonomic richness than the loosely structured bioblitz (e.g., 73 vs. 45 bird species and 104 vs. 63 insect families from the RRSS vs. loosely structured bioblitz, respectively). Models fit to the RRSS data identified seasonal patterns in avian community composition and allowed for estimation of habitat–occupancy relationships for insect taxa. The RRSS protocol has potential for broad transferability as a standardized, quick, and inexpensive way to inventory biodiversity and estimate ecological parameters while providing an outreach opportunity.     

Compliance with and ecosystem function of biodiversity offsets in North American and European freshwaters

Land-use change via human development is a major driver of biodiversity loss. To reduce these impacts, billions of dollars are spent on biodiversity offsets. However, studies evaluating offset project effectiveness that examine components such as the overall compliance and function of projects remain rare. We reviewed 577 offsetting projects in freshwater ecosystems that included the metrics project size, type of aquatic system (e.g., wetland and creek), offsetting measure (e.g., enhancement, restoration, and creation), and an assessment of the projects’ compliance and functional success. Project information was obtained from scientific and government databases and gray literature. Despite considerable investment in offsetting projects, crucial problems persisted. Although compliance and function were related to each other, a high level of compliance did not guarantee a high degree of function. However, large projects relative to area had better function than small projects. Function improved when projects targeted productivity or specific ecosystem features and when multiple complementary management targets were in place. Restorative measures were more likely to achieve targets than creating entirely new ecosystems. Altogether the relationships we found highlight specific ecological processes that may help improve offsetting outcomes.     

Representation of the world's biophysical conditions by the global protected area network

Protected areas (PAs) are often implemented without consideration of already existing PAs, which is likely to cause an overrepresentation of certain biophysical conditions. We assessed the representativeness of the current PA network with regard to the world's biophysical conditions to highlight which conditions are underprotected and where these conditions are located. We overlaid terrestrial and marine PAs with information on biophysical conditions (e.g., temperature, precipitation, and elevation) and then quantified the percentage of area covered by the PA network. For 1 variable at a time in the terrestrial realm, high temperature, low precipitation, and medium and very high elevation were underrepresented. For the marine realm, low and medium sea surface temperature (SST), medium and high sea surface salinity (SSS), and the deep sea were underrepresented. Overall, protection was evenly distributed for elevation across the terrestrial realm and SST across the marine realm. For 2 variables at a time, cold and very dry terrestrial environments had mostly low protection, which was also the case for low SST and low and medium SSS across most depths for marine environments. Low protection occurred mostly in the Sahara and the Arabian Peninsula for the terrestrial realm and along the Tropic of Capricorn and toward the poles for the marine realm. Although biodiversity measures are of prime importance for the design of PA networks, highlighting biophysical gaps in current PAs adds a frequently overlooked perspective. These gaps may weaken the potential of PAs to conserve biodiversity. Thus, our results may provide useful insights for researchers, practitioners, and policy makers to establish a more comprehensive global PA network.     

Quantitative composition of the defensive secretion ofBledius species (Coleoptera: Staphylinidae: Oxytelinae) is adapted to naturally occurring predators

Summary The adaptation of defensive secretions to their target organisms was examined for the abdominal gland secretions ofBledius furcatus, B. spectabilis andB. arenarius. Therefore the target organisms of the secretion of theseBledius species (i.e. their predators) had to be identified. At the collection sites examined these were the earwigLabidura riparia, the antCataglyphis bicolor, the flyLispe candicans, different carabids of the generaPogonus, Dichirotrichus, Dyschirius, Bembidion andCalathus and the wading birdsHaematopus ostralegus andCalidris alba. The secretion of the abdominal glands contains the toxin ptoluquinone dissolved in either-dodecalactone and 1-undecene (B. furcatus andB. spectabilis) or in octanoic acid and octyloctanoate (B. arenarius). The ratio of these solvents is species-specific. Application experiments using some of the natural insect predators (L. riparia, C. bicolor, Pogonus, Di. gustavii, Dyschirius) revealed that these solvent ratios provided a more effective deterrent than other possible ratios. Thus by combining the solvents in certain ratios, the capability of cuticular penetration and therefore the effectiveness of the defensive secretions are adapted to their natural targets.     

Democracy versus development: New urban debates

The politics of development includes subtexts of choice, as underpinned by community consensus (or lack of it), against the larger backdrop of a liberal democracy, with its intricate power structures that influence the choice of concepts and often lead to complicity rather than consent. A dilemma exists between the understanding of field-level practitioners and that of academics, whose understandings of progress differ. Progress can (and often does) imply long-term change across generations whereas development is often more a matter of crisis management and delivering tangible results to end-users. This makes it a bone of contention across political systems, irrespective of local or global dynamics. Development can emerge as a tool to be wielded for power and further political mileage at the expense of progress. Progress cannot be studied as a counterpoint to development as the sense of history is often missing in development interventions. Development tends to be piecemeal as its very logic often stems from the rationale of power. This is not to suggest, however, that development is only about politics; it is definitely more than the politics of everyday life and involves all of civil society that requires its critical and immediate attention. In this sense, it has become a priority for politicians with an underlying agenda designed to out-manouevre dissent and all statements of protest.     

A palaeo-hydrogeological model for arsenic contamination in southern and south-east Asia

An argument is presented in which areas of natural arsenic contamination of modern groundwaters throughout Asia have a common origin. Arsenic originally accumulated in oceanic ferro-manganoan sediments of the eastern Palaeo-Tethys. This was further concentrated through oceanic crustal extinction in what later became the south-east Chinese accreted mineralised terrain. Proto-Himalayan uplift of this area created the palaeo-drainage systems of the Ganges – Brahmaputra, Irrawaddy, Mekong, and Red Rivers, with consequent headwater erosion of arsenic-rich sediments. Their downstream deposition as immature and easily redistributed Neogene sandstones, silts, and iron-rich clays has created secondary and tertiary reservoirs of adsorbed and authigenic arsenic, from which the current arsenic-rich groundwaters have evolved. Considering river basins within the above palaeo-hydrogeological framework provides a basis for assessing the risk of arsenic in groundwater basins of south and south-eastern Asia.     

A framework to measure the wildness of managed large vertebrate populations

As landscapes continue to fall under human influence through habitat loss and fragmentation, fencing is increasingly being used to mitigate anthropogenic threats and enhance the commercial value of wildlife. Subsequent intensification of management potentially erodes wildness by disembodying populations from landscape-level processes, thereby disconnecting species from natural selection. Tools are needed to measure the degree to which populations of large vertebrate species in formally protected and privately owned wildlife areas are self-sustaining and free to adapt. We devised a framework to measure such wildness based on 6 attributes relating to the evolutionary and ecological dynamics of vertebrates (space, disease and parasite resistance, exposure to predation, exposure to limitations and fluctuations of food and water supply, and reproduction). For each attribute, we set empirical, species-specific thresholds between 5 wildness states based on quantifiable management interventions. We analysed data from 205 private wildlife properties with management objectives spanning ecotourism to consumptive utilization to test the framework on 6 herbivore species representing a range of conservation statuses and commercial values. Wildness scores among species differed significantly, and the proportion of populations identified as wild ranged from 12% to 84%, which indicates the tool detected site-scale differences both among populations of different species and populations of the same species under different management regimes. By quantifying wildness, this framework provides practitioners with standardized measurement units that link biodiversity with the sustainable use of wildlife. Applications include informing species management plans at local scales; standardizing the inclusion of managed populations in red-list assessments; and providing a platform for certification and regulation of wildlife-based economies. Applying this framework may help embed wildness as a normative value in policy and mitigate the shifting baseline of what it means to truly conserve a species.     

Sexual Predators,Energy Development,and Conservation in Greater Yellowstone

Abstract: In the United States, as elsewhere, a growing debate pits national energy policy and homeland security against biological conservation. In rural communities the extraction of fossil fuels is often encouraged because of the employment opportunities it offers, although the concomitant itinerant workforce is often associated with increased wildlife poaching. We explored possible positive and negative factors associated with energy extraction in the Greater Yellowstone Ecosystem (GYE), an area known for its national parks, intact biological diversity, and some of the New World's longest terrestrial migrations. Specifically, we asked whether counties with different economies—recreation (ski), agrarian (ranching or farming), and energy extractive (petroleum)—differed in healthcare (gauged by the abundance of hospital beds) and in the frequency of sexual predators. The absolute and relative frequency of registered sex offenders grew approximately two to three times faster in areas reliant on energy extraction. Healthcare among counties did not differ. The strong conflation of community dishevel, as reflected by in‐migrant sexual predators, and ecological decay in Greater Yellowstone is consistent with patterns seen in similar systems from Ecuador to northern Canada, where social and environmental disarray exist around energy boomtowns. In our case, that groups (albeit with different aims) mobilized campaigns to help maintain the quality of rural livelihoods by protecting open space is a positive sign that conservation can matter, especially in the face of rampant and poorly executed energy extraction projects. Our findings further suggest that the public and industry need stronger regulatory action to instill greater vigilance when and where social factors and land conversion impact biological systems.     

Cost‐effective conservation of an endangered frog under uncertainty

How should managers choose among conservation options when resources are scarce and there is uncertainty regarding the effectiveness of actions? Well‐developed tools exist for prioritizing areas for one‐time and binary actions (e.g., protect vs. not protect), but methods for prioritizing incremental or ongoing actions (such as habitat creation and maintenance) remain uncommon. We devised an approach that combines metapopulation viability and cost‐effectiveness analyses to select among alternative conservation actions while accounting for uncertainty. In our study, cost‐effectiveness is the ratio between the benefit of an action and its economic cost, where benefit is the change in metapopulation viability. We applied the approach to the case of the endangered growling grass frog (Litoria raniformis), which is threatened by urban development. We extended a Bayesian model to predict metapopulation viability under 9 urbanization and management scenarios and incorporated the full probability distribution of possible outcomes for each scenario into the cost‐effectiveness analysis. This allowed us to discern between cost‐effective alternatives that were robust to uncertainty and those with a relatively high risk of failure. We found a relatively high risk of extinction following urbanization if the only action was reservation of core habitat; habitat creation actions performed better than enhancement actions; and cost‐effectiveness ranking changed depending on the consideration of uncertainty. Our results suggest that creation and maintenance of wetlands dedicated to L. raniformis is the only cost‐effective action likely to result in a sufficiently low risk of extinction. To our knowledge we are the first study to use Bayesian metapopulation viability analysis to explicitly incorporate parametric and demographic uncertainty into a cost‐effective evaluation of conservation actions. The approach offers guidance to decision makers aiming to achieve cost‐effective conservation under uncertainty.     

Comparison of the insecticidal effects of water extracted and intact aromatic plants onAcanthoscelides obtectus,a bruchid beetle pest of kidney beans

Summary Acanthoscelides obtectus (Say) is one of the most damaging pests of kidney beans,Phaseolus vulgaris L. worldwide. However, aromatic plants from the families Lamiaceae, Lauraceae, Myrtaceae and Poaceae can protect kidney beans by direct or delayed insecticidal effect, through increased adult mortality and inhibition of beetle reproduction (both oviposition and adult emergence). The efficiency of hydrodistillated and intact plants fromThymus vulgaris andT. serpyllum, Mentha piperata, Rosmarinus officinalis, Satureia hortensis, Eucalyptus globulus, Laurus nobilis, Origanum vulgare, andCymbopogon nardus was compared. For both extracts,Origanum vulgare had the best effect. The insecticidal effect was induced by more than the essential oils because no significant difference was noticed between distilled and intact plants extract. Inhibition of reproduction was particularly important. These results suggest that lipidic as well as non lipidic, allelochemicals, such as phenolics, or non-protein amino-acids, or flavonoids may be involved in the toxicity of aromatic plants to this beetle.