Biosynthesis of citric acid by Yarrowia lipolytica repeat-batch culture on ethanol

After analysis of batch culture and identification of the ways for prolongation of citric acid active synthesis by yeast, repeat-batch (RB) cultivation was suggested. Yarrowia lipolytica strain RB cultivation was studied and optimal conditions for cultivation selected. It was shown that when applying RB cultivation, better results were obtained than for batch cultivation. The activity of the culture remained stable after cultivation for more than 700 h. Comparative analysis of enzyme activities confirmed the regularity of the effect described, as the activity of practically of all the enzymes participating in ethanol oxidation and citric acid biosynthesis remained stable over time during RB cultivation. Advantages of RB cultivation for the production of citric acid by yeast are discussed. Received: 1 March 1999 / Received revision: 28 June 1999 / Accepted: 5 July 1999     

A two-stage cultivation process for the growth enhancement of Chlorella vulgaris

This study proposes a two-stage cultivation process with an autotrophic growth followed by a mixotrophic process. The results indicated that a two-stage cultivation process using a daily dose of 3 g/L of glucose could achieve 7.4 g/L of biomass, which was about a 64 % increase over simple autotrophic cultivation. In the second stage of mixotrophic cultivation, glucose was regarded as a better carbon source for cell growth, than was glycerol. Linoleic acid (C18:2) would be the primary component in the two-stage cultivation as in the autotrophic cultivation. Even carbon source was provided in the second stage of mixotrophic cultivation; lower light intensity limited the mixotrophic growth, which indicated that photosynthesis still plays an important role in the second stage of mixotrophical cultivation. The final biomass was higher after this two-stage cultivation process, which made it suitable for application in the production scale-up of algal biomass.     

Biosynthesis of amylolytic enzymes during continous cultivation of the yeast-like fungus Endomycopsis fibuliger 55-13

Growth of the yeast-like fungus Endomycopsis fibuliger 55-13 during continous cultivation was studied. The microorganism produced amylolytic enzymes during flow cultivation. As a result of the three-stage cultivation the productivity increased two-fold and the time of cultivation decreased by 6 hrs as compared with periodic cultivation.     

Kinetics of continuous cultivation

Kinetic, studies were made on continuous cultivation applying the theory of microbial cell growth that was derived previously by the authors introducing the concepts of critical concentration and coefficient of consumption activity. General equations for microbial cell concentration for continuous cultivation in continuous-stirred tank and tubular type reactors were derived theoretically. Productivity of cell mass in continuous cultivation was analyzed kinetically and the behavior of mutant populations in continuous cultivation is briefly discussed.     

杏鲍菇菌丝体水溶性多糖提取及培养条件优化

以马铃薯葡萄糖综合培养基(PDP)为基础培养基,采用正交试验法优化杏鲍菇菌丝体多糖发酵条件,对接种量、摇床转速和培养时间等因素对多糖含量的影响进行了研究。采用水提醇沉法提取多糖,苯酚—浓硫酸法进行多糖含量测定。结果表明,最佳培养条件:接种量为每瓶1块直径为1cm的菌块,转速为140r/min,培养时间为8d。此时杏鲍菇菌丝体多糖含量最高,为75.1mg/g。     

Dynamics of Shifting Cultivation Landscapes in Northern Lao PDR Between 2000 and 2009 Based on an Analysis of MODIS Time Series and Landsat Images

The rotational nature of shifting cultivation poses several challenges to its detection by remote sensing. Consequently, there is a lack of spatial data on the dynamics of shifting cultivation landscapes on a regional, i.e. sub-national, or national level. We present an approach based on a time series of Landsat and MODIS data and landscape metrics to delineate the dynamics of shifting cultivation landscapes. Our results reveal that shifting cultivation is a land use system still widely and dynamically utilized in northern Laos. While there is an overall reduction in the areas dominated by shifting cultivation, some regions also show an expansion. A review of relevant reports and articles indicates that policies tend to lead to a reduction while market forces can result in both expansion and reduction. For a better understanding of the different factors affecting shifting cultivation landscapes in Laos, further research should focus on spatially explicit analyses.     

嫁接与施氮对甜瓜产量和氮素吸收、利用的影响

通过裂区设计田间试验,主区为2种栽培方式(嫁接栽培和自根栽培),副区为4个施氮水平(0、120、240、360 kg N·hm^-2),研究了栽培方式和施氮量对甜瓜产量和品质、氮素运移和分配,以及氮素利用率的影响.结果表明: 嫁接栽培的甜瓜商品瓜产量较自根甜瓜提高了7.3%,可溶性固形物含量降低了0.16%～3.28%;生长前期嫁接栽培甜瓜氮素累积量较自根栽培低,结果后嫁接栽培氮素累积量显著升高,收获时植株氮素累积量较自根栽培增加了5.2%,果实中的氮素累积量提高了10.3%;嫁接栽培植株氮素向果实的转移量较自根栽培提高了20.9%,嫁接栽培果实中的氮素分配率在80%以上,自根栽培的分配率在80%以下;在同一施氮水平下,嫁接栽培的甜瓜氮素吸收利用率较自根栽培提高了1.3%～4.2%,氮素农学效率提高了2.73～5.56 kg·kg^-1,氮素生理利用率提高了7.39～16.18 kg·kg^-1;从商品瓜产量、氮素吸收量和氮素利用率综合考虑,施氮量240 kg·hm^-2为本区域嫁接甜瓜较适宜的氮素用量.     

Fed-batch cultivation for the production of clavulanic acid by an immobilized Streptomyces clavuligerus mutant

In order to obtain high productivity of clavulanic acid, a newly-introduced carrier, polyurethane pellet (PUP) Z97-020 was used for the immobilization process. In a stirred-tank bioreactor, batch cultivation by Streptomyces clavuligerus KK immobilized on PUP Z97-020 gave about 3100 mg of clavulanic acid per litre, representing an increase of 200% in productivity compared with that by fed-batch cultivation of free cells (1500 mg/l). However, the clavulanic acid produced rapidly decomposed due to the pH change during batch cultivation. Fed-batch cultivation by immobilized S. clavuligerus KK gave an excellent level of clavulanic acid up to 3250 mg/l, a productivity increase of 220% compared with that by fed-batch cultivation of free cells. These results suggest that immobilization with PUP Z97-020 is a more effective process for the production of clavulanic acid and that the maintenance of pH by fed-batch cultivation with glycerol as a limiting substrate prevents the clavulanic acid from decomposing during the fermentation.     

Biocatalytic decolourisation of molasses by Phanerochaete chrysosporium

Bioremediation potential of Phanerochaete chrysosporium strains NCIM 1073, NCIM 1106 and NCIM 1197 to decolourise molasses in solid and liquid molasses media was studied. Strains varied in the pattern of molasses decolourisation on solid medium by Giant colony method. Under submerged cultivation conditions, strain NCIM 1073 did not decolourise molasses while, strains NCIM 1106 and NCIM 1197 could decolourise molasses up to 82% and 76%, respectively. Under stationary cultivation conditions, none of the strains could decolourise molasses. This was overcome by increasing the surface area of the culture in flat bottom glass bottles under stationary cultivation conditions. Under submerged cultivation conditions, growth was more or less same in all strains. However, the lignin peroxidase and manganese peroxidase activities were significantly less in the strain NCIM 1073. Under stationary cultivation conditions, none of the strains could produce enzymes lignin peroxidase, manganese peroxidase and laccase. However, all of them could produce lignin peroxidase and manganese peroxidase when cultivated in flat bottom glass bottles under stationary cultivation conditions.     

Application of terminal restriction fragment length polymorphism (T-RFLP) analysis to monitor effect of biocontrol agents on rhizosphere microbial community of hot pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.)

Microbial communities in hot pepper (Capsicum annuum L.) cultivation fields under different cultivation methods were investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis. Rhizosphere soil and leaf samples were collected from control, conventional and nature-friendly cultivation fields between May and July, 2009. Two Bacillus subtilis strains were applied to nature-friendly cultivation fields as biocontrol agents during the sampling period. Relative abundances of bacteria and plant pathogenic fungi related T-RFs were also measured to monitor the effect of biocontrol agents on potential plant pathogenic fungi. In the principal component analysis (PCA) based on T-RFLP profiles, the microbial communities from rhizosphere soil samples in July, including bacteria and fungi, showed distinct difference between nature-friendly cultivation fields and other cultivation fields. However, there was no correlation between cultivation methods and leaf microbial communities at any sampling period. Changes in the abundance of bacteria related T-RF in the rhizosphere of nature-friendly cultivation fields were observed clearly two months after application of biocontrol agent, while the abundance of plant pathogenic fungi related T-RFs significantly decreased.     

Simultaneous measurement of ethylene and CO2 production and of O2 consumption in soil and pure microbial cultures

Summary The method described here makes it possible to determine gaseous and volatile compounds produced by stationary and shaken cultures of microorganisms, and by soil samples. In a closed system oxygen consumed by a biological sample is electrolytically replenished and CO₂ produced is trapped in a KOH solution. Oxygen from the electrolyzer was replenished through a distribution bottle with the aid of a system of polyethylene tubings equipped with injection needles at both ends. The amount of the oxygen consumed is permanently reflected by volume of the electrolytically released hydrogen. The reverse flow of atmosphere from the cultivation flasks was prevented by a liquid seal formed by a KOH solution or the cultivation medium itself. Suba-seal type flasks (125 ml) with rubber caps served for the cultivation. Glass cylinders placed inside cultivation flasks or penicillin flasks (20 ml) connected with cultivation flasks by polyethylene tubings served as adsorption vessels. By exchanging the KOH solution it is possible to follow CO₂ production even during individual cultivation phases. Volatile acids adsorbed in the KOH solution could be determined after their release by mineral acid. Samples of atmosphere were analyzed by gas chromatography. The whole system had to be temperature equilibrated.     

Hopped Beer: The Case For Cultivation

The history of hops, hopped beer, and hop cultivation is unclear and ambiguous. An assessment of the available literature reveals many contradictions, especially regarding the first use of hops in beer and the earliest incidence of hop cultivation. Historically, hops were used for a variety of purposes; now their primary use is as a preservative and flavoring in beer. Hop cultivation is poorly documented, but was certainly undertaken by the 10th century, most probably in response to the demand generated by beer-brewing. After comparing the literature and investigating source material, a chronology of hop use in beer and hop cultivation is proposed.     

Socioeconomic determinants of cultivation of non-wood forest products in southern Nigeria

Cultivation of non-wood forest products (NWFP), although a veritable means of ecosystem and biodiversity conservation and improved natural resource management, has not been sustained in southern Nigeria (Nigeria’s major forest region) notwithstanding the unprecedented rate of depletion of the resource in the wild. For example, efforts in the past to support cultivation of NWFP in southern Nigeria under a USAID funded Cross River State forestry project, especially through nursery establishment for some rural communities, was not sustained as the initiative was abandoned by the participants. Hence to promote cultivation of NWFP, this study ascertained socioeconomic factors that influence cultivation of NWFPs. Multi stage sampling technique was used in selection of respondents (households) from two States: Cross River and Enugu States of southern Nigeria. A sample size of 400 households was used for the study. Probit and multinomial logistic regressions were used in estimating NWFP cultivation. The findings show that cultivation of NWFP was positively determined by gender, farming occupation (especially of women farmers) distance to forests where NWFP was collected, proportion of household food from NWFP, and medium wealth category. Age had a negative effect on cultivation until the age of 46 after which the effect became positive. In addition, household size, gender and farming occupation of household heads positively influenced cultivation of NWFP in plantations as against home gardens while gender and farming occupation had positive effect on cultivation of some stands of NWFP as against home gardens. Incorporating the findings of this study in future intervention project for NWFP cultivation will help sustain the initiative.     

The optimization of the batch cultivation process for Pseudomonas aeruginosa to produce exotoxin A

The experimental study of conditions for the optimization of the batch cultivation of P. aeruginosa has been made. As revealed in this study the aim of this cultivation can be achieved by using exponentially growing culture in a dose of 1.10(9) cells/ml as seed material and by ensuring the conditions of rational air supply during the main cultivation process.     

恶臭假单胞菌NA-1菌体培养转化和静息细胞转化联合工艺生产6-羟基烟酸研究

现有微生物羟基化烟酸采用的是静息细胞转化工艺。但研究揭示,恶臭假单胞菌NA-1(Pseudomonas putidaNA-1)在培养过程中不降解发酵液中由诱导剂烟酸转化形成的6-羟基烟酸,这是由于烟酸的存在抑制了羟基烟酸降解酶的作用,而不是因为细胞停止生长不利用羟基烟酸的缘故。因而尝试利用菌体诱导培养过程进行烟酸转化生产,建立了一种新的生产工艺,即菌体培养转化和静息细胞转化联合工艺。该工艺在恶臭假单胞菌NA-1培养过程中持续补充烟酸以维持1%(W/V)浓度,使烟酸被生长细胞转化为羟基化烟酸并在发酵液中线性积累,而不被进一步降解;培养转化结束后,发酵液中的静息细胞依然拥有很高的羟基化酶活力,能够再次用于转化反应。该联合转化工艺与传统的静息细胞转化工艺相比,不仅节约了诱导剂烟酸,而且6-羟基烟酸的产量提高了65%。     

Synthesis of alpha-fetoprotein, albumin and transferrin by long-term cultured cells of murine hepatoma]

The capacity of continuous cell of XXIIa mouse hepatoma (strain MHXXIIa) to synthesize alpha-fetoprotein, albumin and transferrin was studied by immunoautoradiography. Albumin and transferrin were detected in the polyethylene glycol concentrated growth medium of hepatoma cells on the 5th year (the 55th month) of their cultivation. alpha-fetoprotein was not found. Only transferrin was revealed in the growth medium of hepa toma cells of the 8th year (the 92d month) of cultivation. Two clonal cultures obtained on the 8th year of hepatoma cell cultivation were also characterized by the ability to synthesize transferrin. The continuous mouse hepatoma cells retained their malignancy. The agar micro-precipitation reaction showed the presence of alpha-fetofetoprotein in lyfogel concentrated serum of mice with tumors formed after inoculation of the hepatoma cells of the 5th year of cultivation. However, alpha-fetoprotein was not detected in the serum of mice with tumors induced by inoculation of the hepatoma cells of the 8th year of cultivation.     

Change in the composition of a bacterial association degrading aromatic compounds during oil sludge detoxification in a continuous-flow microbial reactor

Analysis of oil sludge by direct plating and enrichment cultivation revealed 16 strains degrading aromatic compounds. After 30 days of cultivation in a continuous-flow microbial reactor, 17 more degrader strains were isolated. Genotyping of these strains showed that they were taxonomically diverse, and the range of strains degrading naphthalene, benzene, toluene, ethylbenzene, and xylenes depended on isolation methods. Direct plating yielded more aromatic degraders than enrichment cultivation. A microbial association different from that existing before the enrichment cultivation was obtained in the laboratory continuous-flow reactor.     

不同荫蔽栽培下亏缺灌溉对干热区小粒咖啡水光利用和产量的影响

干热区小粒咖啡水光管理粗放,产量和品质得不到保证.为探明干热区小粒咖啡最佳灌水和荫蔽栽培耦合模式,通过大田试验,设3个灌水水平(充分灌水、轻度亏缺灌水和重度亏缺灌水)和4个荫蔽栽培模式(无荫蔽:单作咖啡;轻度荫蔽:4行咖啡间作1行香蕉;中度荫蔽:3行咖啡间作1行香蕉;重度荫蔽:2行咖啡间作1行香蕉),研究香蕉荫蔽栽培下亏缺灌溉对小粒咖啡生长、叶片光合特性、水光利用和产量的影响.结果表明: 小粒咖啡叶片的净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(g_s)、叶片水分利用效率(LWUE)、叶片表观光能利用效率(LRUE)随灌水量的增大而增大,胞间CO₂浓度(C_i)随灌水量的增大而减小;与充分灌水相比,轻度亏缺灌水的干豆产量减小9.4%,重度亏缺灌水的干豆产量减小36.7%,水分利用效率(WUE)减小16.9%.P_n、T_r、g_s、LWUE随荫蔽度的增大呈先增大后减小的趋势,中度荫蔽栽培的增量最大;与无荫蔽模式相比,轻度荫蔽模式干豆产量增加13.0%,WUE增加12.9%,中度荫蔽栽培模式干豆产量增加23.1%,WUE增加23.4%.干豆产量、WUE、百粒咖啡豆的体积和百粒鲜质量随灌水量和荫蔽度的增大呈不同程度增大,其中,中度荫蔽栽培下充分灌水的干豆产量和WUE增量最大.相同土层深度的土壤含水率随荫蔽度的增加而减小;在0~50 cm土层,土壤含水率随土层深度的增加先增大后减小.LRUE与光合有效辐射呈显著的负指数关系或符合Logistic曲线变化.因此,从优质高产、水光高效利用的综合效益考虑,中度荫蔽栽培下充分灌水是小粒咖啡灌水处理和香蕉荫蔽栽培模式的最佳组合.     

Untangling a Decline in Tropical Forest Resilience: Constraints on the Sustainability of Shifting Cultivation Across the Globe

Shifting cultivators depend on forest biomass inputs to nourish their crops. For them, forest resilience has an immediate impact: it affects crop productivity. A decline in the rate of recovery following shifting cultivation would ultimately affect local, regional and global carbon budgets, with feedbacks to climate. Yet the long-term impacts of shifting cultivation have been quantified in only six locations. In this study, we reanalyze data from these locations to determine whether the rate of biomass recovery is the same from cycle to cycle. Further, using case studies in Southern Yucatan, Mexico and West Kalimantan, Indonesia, we investigate the ecological and socioeconomic factors that affect forest resilience and thus determine whether or not shifting cultivation is sustainable. The reanalysis links aboveground biomass recovery following shifting cultivation to site productivity, forest age, fallow length, history of cultivation, and soil texture. Across locations, biomass accumulation rate declines by 9.3 percent with each cycle of shifting cultivation. Per cycle change in biomass accumulation rate is significantly more negative in younger forests and forests that experience a shorter fallow period. However, more detailed analyses for two case studies suggest that a purely ecological framework is of limited effectiveness in explaining variability in the effect of repeated shifting cultivation. Rather, socioeconomic factors such as migration, subsidies, roads, and settlement history can alter the outcome of shifting cultivation by limiting the accumulation and use of local knowledge.     

Change in the composition of a bacterial association degrading aromatic compounds during oil sludge detoxification in a continuous-flow microbial reactor

Analysis of oil sludge by direct plating and enrichment cultivation revealed 16 strains degrading aromatic compounds. After 30 days of cultivation in a continuous-flow microbial reactor, 17 more degrader strains were isolated. Genotyping of these strains showed that they were taxonomically diverse, and the range of strains degrading naphthalene, benzene, toluene, ethylbenzene, and xylenes depended on isolation methods. Direct plating yielded more aromatic degraders than enrichment cultivation. A microbial association different from that existing before the enrichment cultivation was obtained in the laboratory continuous-flow reactor.