Showing papers in "Journal of Industrial Microbiology & Biotechnology in 1996"

Biodiversity and application of microalgae

Abstract: The algae are a polyphyletic, artificial assemblage of O2-evolving, photosynthetic organisms (and secondarily nonphotosynthetic evolutionary descendants) that includes seaweeds (macroalgae) and a highly diverse group of microorganisms known as microalgae. Phycology, the study of algae, developed historically as a discipline focused on the morphological, physiological and ecological similarities of the subject organisms, including the prokaryotic bluegreen algae (cyanobacteria) and prochlorophytes. Eukaryotic algal groups represent at least five distinct evolutionary lineages, some of which include protists traditionally recognized as fungi and protozoa. Ubiquitous in marine, freshwater and terrestrial habitats and possessing broad biochemical diversity, the number of algal species has been estimated at between one and ten million, most of which are microalgae. The implied biochemical diversity is the basis for many biotechnological and industrial applications.

Total bacterial diversity in soil and sediment communities—A review

Abstract: Advances in microbial methods have demonstrated that microorganisms globally are the dominating organisms both concerning biomass and diversity Their functional and genetic potential may exceed that of higher organisms Studies of bacterial diversity have been hampered by their dependence on phenotypic characterization of bacterial isolates Molecular techniques have provided the tools for analyzing the entire bacterial community including those which we are not able to grow in the laboratory Reassociation analysis of DNA isolated directly from the bacteria in pristine soil and marine sediment samples revealed that such environments contained in the order of 10 000 bacterial types The diversity of the total bacterial community was approximately 170 times higher than the diversity of the collection of bacterial isolates from the same soil The culturing conditions therefore select for a small and probably skewed fraction of the organisms present in the environment Environmental stress and agricultural management reduce the bacterial diversity With the reassociation technique it was demonstrated that in heavily polluted fish farm sediments the diversity was reduced by a factor of 200 as compared to pristine sediments Here we discuss some molecular mechanisms and environmental factors controlling the bacterial diversity in soil and sediments

Environmental applications of immobilized microbial cells: A review

Abstract: Immobilized microbial cells have been used extensively in various industrial and scientific endeavours. However, immobilized cells have not been used widely for environmental applications. This review examines many of the scientific and technical aspects involved in using immobilized microbial cells in environmental applications, with a particular focus on cells encapsulated in biopolymer gels. Some advantages and limitations of using immobilized cells in bioreactor studies are also discussed.

Bioactive compounds produced by cyanobacteria

Abstract: Cyanobacteria produce a large number of compounds with varying bioactivities. Prominent among these are toxins: hepatotoxins such as microcystins and nodularins and neurotoxins such as anatoxins and saxitoxins. Cytotoxicity to tumor cells has been demonstrated for other cyanobacterial products, including 9-deazaadenosine, dolastatin 13 and analogs. A number of compounds in cyanobacteria are inhibitors of proteases — micropeptins, cyanopeptolins, oscillapeptin, microviridin, aeruginosins- and other enzymes, while still other compounds have no recognized biological activities. In general cyclic peptides and depsipeptides are the most common structural types, but a wide variety of other types are also found: linear peptides, guanidines, phosphonates, purines and macrolides. The close similarity or identity in structures between cyanobacterial products and compounds isolated from sponges, tunicates and other marine invertebrates suggests the latter compounds may be derived from dietary or symbiotic blue-green algae.

Secondary metabolites of the fungus Monascus : A review

Abstract: This review deals with polyketides produced by the filamentous fungusMonascus which include: 1) a group of yellow, orange and red pigments, 2) a group of antihypercholesterolemic agents including mevinolin and related compounds and 3) the newly discovered metabolite ankalactone. Biosynthesis, methods of production, isolation and biological activities of these secondary metabolites are discussed.

Review: optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter.

Abstract: This review examines factors which influence the expression of foreign proteins inEscherichia coli under the transcriptional control of thelac andtac promoters, and discusses conditions for maximizing the production of a foreign protein using this system. Specifically, the influence of IPTG (isopropyl-β-d-thiogalactoside) concentration, temperature, composition of the growth medium, the point in the growth curve at which cells are induced with either IPTG or lactose, and the duration of the induction phase are discussed.

Quantitative comparisons of in situ microbial biodiversity by signature biomarker analysis

Abstract: Microscopic examinations have convinced microbial ecologists that the culturable microbes recovered from environmental samples represent a tiny proportion of the extant microbiota. Methods for recovery and enzymatic amplification of nucleic acids from environmental samples have shown that a huge diversity existsin situ, far exceeding any expectations which were based on direct microscopy. It is now theoretically possible to extract, amplify and sequence all the nucleic acids from a community and thereby gain a comprehensive measure of the diversity as well as some insights into the phylogeny of the various elements within this community. Unfortunately, this analysis becomes economically prohibitive if applied to the multitude of niches in a single biome let alone to a diverse set of environments. It is also difficult to utilize PCR amplification on nucleic acids from some biomes because of coextracting enzymatic inhibitors. Signature biomarker analysis which potentially combines gene probe and lipid analysis on the same sample, can serve as a complement to massive environmental genome analysis in providing quantitative comparisons between microniches in the biome under study. This analysis can also give indications of the magnitude of differences in biodiversity in the blome as well as provide insight into the phenotypic activities of each community in a rapid and cost-effective manner. Applications of signature lipid biomarker analysis to define quantitatively the microbial viable biomass of portions of an Eastern USA deciduous forest, are presented.

Diverse reactions catalyzed by naphthalene dioxygenase from Pseudomonas sp strain NCIB 9816

Abstract: Naphthalene dioxygenase (NDO) fromPseudomonas sp strain NCIB 9816 is a multicomponent enzyme system which initiates naphthalene catabolism by catalyzing the addition of both atoms of molecular oxygen and two hydrogen atoms to the substrate to yield enantiomerically pure (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene. NDO has a relaxed substrate specificity and catalyzes the dioxygenation of many related 2- and 3-ring aromatic and hydroaromatic (benzocyclic) compounds to their respectivecis-diols. Biotransformations with a diol-accumulating mutant, recombinant strains and purified enzyme components have established that in addition tocis-dihydroxylation, NDO also catalyzes a variety of other oxidations which include monohydroxylation, desaturation (dehydrogenation),O-andN-dealkylation and sulfoxidation reactions. In several cases, the absolute stereochemistry of the oxidation products formed by NDO are opposite to those formed by toluene dioxygenase (TDO). The reactions catalyzed by NDO and other microbial dioxygenases can yield specific hydroxylated compounds which can serve as chiral synthons in the preparation of a variety of compounds of interest to pharmaceutical and specialty chemical industries. We present here recent work documenting the diverse array of oxidation reactions catalyzed by NDO. The trends observed in the oxidation of a series of benzocyclic aromatic compounds are compared to those observed with TDO and provide the basis for prediction of regio- and stereospecificity in the oxidation of related substrates. Based on the types of reactions catalyzed and the biochemical characteristics of NDO, a mechanism for oxygen activation by NDO is proposed.

Cyclic peptides and depsipeptides from cyanobacteria: a review.

Abstract: An elaborate array of structurally-novel and biologically-active cyclic peptides and depsipeptides are found in blue-green algae (cyanobacteria). Several of these compounds possess structures that are similar to those of natural products from marine invertebrates. Most of these cyclic peptides and depsipeptides, such as the microcystins and the lyngbyatoxins, will probably only be useful as biochemical research tools. A few, however, have the potential for development into useful commercial products. For example, cryptophycin-1, a novel inhibitor of microtubule assembly from Nostoc sp GSV 224, shows impressive activity against a broad spectrum of solid tumors implanted in mice, including multidrug-resistant ones, and majusculamide C, a microfilament-depolymerizing agent from Lyngbya majuscula, shows potent fungicidal activity and may have use in the treatment of resistant fungal-induced diseases of domestic plants and agricultural crops.

Taxol from fungal endophytes and the issue of biodiversity

Abstract: Fungi represent one of the most understudied and diverse group of organisms. Commonly, these organisms make associations with higher life forms and may proceed to biochemically mimic the host organism. An excellent example of this is the anticancer drug, taxol, which had been previously supposed to occur only in the plant genusTaxus (yew). However, taxol has been reported in a novel endophytic fungus—Taxomyces andreanae, but also has been demonstrated to occur in a number of unrelated fungal endophytes includingPestalotia, Pestalotiopsis, Fusarium, Alternaria, Pithomyces, Monochaetia and others. Thus, this report presents information on the presence of taxol among disparate fungal genera, and uses these observations as an additional argument to support efforts to study fungal endophytes and preserve their associated host plants.

Identification of an anaerobic bacterium which reduces perchlorate and chlorate as Wolinella succinogenes

Abstract: A bacterium coded as strain HAP-1 was isolated from a municipal anaerobic digestor for its ability to reduce >7000 ppm perchlorate in wastewaters. The organism is capable of the dissimilatory reduction of perchlorate on chlorate to chloride for energy and growth. It is a Gram-negative, non-sporeforming, obligately anaerobic, motile thin rod. Antibiotic resistance, utilization of carbon substrates and utilization of electron acceptors by bacterium HAP-1 were similar toWolinella succinogenes. The organism's 16S rRNA sequence was 0.75% different from that of the type strain ofW. succinogenes. The fatty acid compositions of the two organisms are very similar. The morphological, physiological and 16S rRNA sequence data indicated that bacterium HAP-1 is a strain ofW. succinogenes that can utilize perchlorate or chlorate as a terminal electron acceptor.

Genetic identification of biological species in the Saccharomyces sensu stricto complex

Abstract: Studies on taxonomic and evolutionary genetics of theSaccharomyces sensu stricto complex are considered in light of the biological species concept. Genetic variability of some physiological properties traditionally used in yeast taxonomy is discussed. Genetic hybridization analysis and molecular karyotyping revealed six biological species in theSaccharomyces sensu stricto complex. DNA-DNA reassociation data are concordant with the data obtained by genetic analysis. A new system for naming the cultivatedSaccharomyces yeast (groups of cultivars) is proposed.

Biodiversity of freshwater fungi

Abstract: There are more than 600 species of freshwater fungi with more known from temperate, as compared to tropical regions. These includeca 340 ascomycetes, 300 deuteromycetes, and a number of lower fungi which are not discussed here.Aniptodera, Annulatascus, Massarina, Ophioceras andPseudohalonectria are common freshwater ascomycetes, which appear to be well adapted for this lifestyle either in their ascospore types or their competitive-degradative characters. The most common genera of wood-inhabiting deuteromycetes includeCancellidium, Dactylaria, Dictyosporium andHelicomyces. They are categorized into four groups depending on their form and life style: the ingoldian hyphomycetes; the aero-aquatic hyphomycetes; the terrestrial-aquatic hyphomycetes; and the submerged-aquatic hyphomycetes. The adaptations of aquatic fungi for their dispersal and subsequent attachment to new substrates are discussed.

Tracing the interaction of bacteriophage with bacterial biofilms using fluorescent and chromogenic probes

Abstract: Phages T4 and E79 were fluorescently-labeled with rhodamine isothiocyanate (RITC), fluoroscein isothiocyanate (FITC), and by the addition of 4'6-diamidino-2-phenylindole (DAPI) to phage-infected host cells of Escherichia coli and Pseudomonas aeruginosa. Comparisons of electron micrographs with scanning confocal laser microscope (SCLM) images indicated that single RITC-labeled phage particles could be visualized. Biofilms of each bacterium were infected by labeled phage. SCLM and epifluorescence microscopy were used to observe adsorption of phage to single-layer surface-attached bacteria and thicker biofilms. The spread of the recombinant T4 phage, YZA1 (containing an rII-LacZ fusion), within a lac E. coli biofilm could be detected in the presence of chromogenic and fluorogenic homologs of galactose. Infected cells exhibited blue pigmentation and fluorescence from the cleavage products produced by the phage-encoded beta-galactosidase activity. Fluorescent antibodies were used to detect non-labeled progeny phage. Phage T4 infected both surface-attached and surface-associated E. coli while phage E79 adsorbed to P. aeruginosa cells on the surface of the biofilm, but access to cells deep in biofilms was somewhat restricted. Temperature and nutrient concentration did not affect susceptibility to phage infection, but lower temperature and low nutrients extended the time-to-lysis and slowed the spread of infection within the biofilm.

Control of microbial souring by nitrate, nitrite or glutaraldehyde injection in a sandstone column

Abstract: Microbial souring (production of hydrogen sulfide by sulfate-reducing bacteria, SRB) in crushed Berea sandstone columns with oil field-produced water consortia incubated at 60°C was inhibited by the addition of nitrate (NO3) or nitrite (NO 2 − ). Added nitrate (as nitrogen) at a concentration of 0.71 mM resulted in the production of 0.57–0.71 mM nitrite by the native microbial population present during souring and suppressed sulfate reduction to below detection limits. Nitrate added at 0.36 mM did not inhibit active souring but was enough to maintain inhibition if the column had been previously treated with 0.71 mM or greater. Continuous addition of 0.71–0.86 mM nitrite also completely inhibited souring in the column. Pulses of nitrite were more effective than the same amount of nitrite added continuously. Nitrite was more effective at inhibiting souring than was glutaraldehyde, and SRB recovery was delayed longer with nitrite than with glutaraldehyde. It was hypothesized that glutaraldehyde killed SRB while nitrite provided a long-term inhibition without cell death. Removal of nitrate after as long as 3 months of continuous addition allowed SRB in a biofilm to return to their previous level of activity. Inhibition was achieved with much lower levels of nitrate and nitrite, and at higher temperatures, than noted by other researchers.

Biodiversity of microorganisms that degrade bacterial and synthetic polyesters.

Abstract: The biodiversity and occurrence in nature of bioplastic-degrading microorganisms are exemplified by the identification of 695 strains, isolated from different environments, such as soils, composts, natural waters, and sludge, that are able to degrade the bacterial polyester poly(3-hydroxybutyrate)in vitro. These microorganisms belong to at least 57 different taxa, including Gram-negative and Gram-positive bacteria, streptomycetes, and moulds. The literature on the biodiversity of poly(3-hydroxybutyrate)-degrading microorganisms is reviewed. The degrading abilities of 171 streptomycete strains were investigated on four different bacterial poly(3-hydroxyalkanoates), and the synthetic polyesters poly(e-caprolactone) and BIONOLLE, and most of these strains degraded at least three different polymers.

A direct comparison of approaches for increasing carbon flow to aromatic biosynthesis in Escherichia coli

Abstract: Different approaches to increasing carbon commitment to aromatic amino acid biosynthesis were compared in isogenic strains ofEscherichia coli. In a strain having a wild-type PEP: glucose phosphotransferase (PTS) system, inactivation of the genes encoding pyruvate kinase (pykA andpykF) resulted in a 3.4-fold increase in carbon flow to aromatic biosynthesis. In a strain already having increased carbon flow to aromatics by virtue of overexpression of thetktA gene (encoding transketolase), thepykA and/orpykF mutations had no effect. A PTS glucose+ mutant showed a 1.6-fold increase in carbon flow to aromatics compared to the PTS+ control strain. In the PTS− glucose+ host background, overexpression oftktA caused a further 3.7-fold increase in carbon flow, while inactivation ofpykA andpykF caused a 5.8-fold increase. When all of the variables tested (PTS− glucose+,pykA, pykF, and overexpressedtktA) were combined in a single strain, a 19.9-fold increase in carbon commitment to aromatic biosynthesis was achieved.

A molecular approach to search for diversity among bacteria in the environment

Abstract: Molecular 16S rDNA-based techniques were applied to a peat sample from northern Germany in order to investigate the bacterial diversity present and compare the clone sequences with those obtained from similar studies on other terrestrial samples. Genomic DNA was extracted from the peat matrix by a direct lysis procedure. 16S rRNA genes were amplified using PCR primers targeting conserved regions of bacterial 16S rDNA. 16S rDNA fragments were blunt end cloned into a plasmid vector and the resulting clone library of 262 sequences was screened by hybridization with different oligonucleotide probes and sequence analysis of randomly selected clones. The 16S rDNA insert of 76 clones was partially sequenced. Clones identified either by hybridization or by sequence analysis fell into three phyla. As judged by hybridization with a specific oligonucleotide probe, 42% of the clones represented members of the alpha subclass of Proteobacteria. Twenty-five of these clones were selected randomly for sequence analysis; none could be assigned to any of the known genera of this subclass. The second largest clone group comprises 15% of the clones and clusters aroundAcidimicrobium ferrooxidans andRubrobacter radiotolerans, both of which are remotely related to members of the order Actinomycetales. The third major clone cluster (10%) was moderately to remotely related to theAcidobacterium capsulatum phylum. Of the additional clones sequenced, a few could be assigned to other subclasses ofProteobacteria, theVerrucomicrobium phylum and the phylum of spirochetes. Comparison of the results presented here with those from other environments reveals a significant number of common clone clusters. As the vast majority of sequences retrieved from any of the marine and terrestrial samples investigated so far by molecular methods indicate the presence of novel bacterial species it can be assumed that a huge, as yet untapped biotechnological potential is present in the environment.

Ethanol production from hemicellulose hydrolysates of agricultural residues using genetically engineered Escherichia coli strain KO11.

Abstract: Hemicellulose hydrolysates of the agricultural residues bagasse, corn stover, and corn hulls plus fibers were readily fermented to ethanol by recombinantEscherichia coli strain KO11. Corn steep liquor and crude yeast autolysate served as excellent nutrients. Fermentations were substantially complete within 48 h, often achieving over 40 g ethanol L−1. Ethanol yields ranged from 86% to over 100% of the maximum theoretical yield (0.51 g ethanol g sugar−1.

Measurement of hydrocarbon-degrading microbial populations by a 96-well plate most-probable-number procedure

Abstract: A 96-well microtiter plate most-probable-number (MPN) procedure was developed to enumerate hydrocarbon-degrading microorganisms. The performance of this method, which uses number 2 fuel oil (F2) as the selective growth substrate and reduction of iodonitrotetrazolium violet (INT) to detect positive wells, was evaluated by comparison with an established 24-well microtiter plate MPN procedure (the Sheen Screen), which uses weathered North Slope crude oil as the selective substrate and detects positive wells by emulsification or dispersion of the oil. Both procedures gave similar estimates of the hydrocarbon-degrader population densities in several oil-degrading enrichment cultures and sand samples from a variety of coastal sites. Although several oils were effective substrates for the 96-well procedure, the combination of F2 with INT was best, because the color change associated with INT reduction was more easily detected in the small wells than was disruption of the crude oil slick. The method's accuracy was evaluated by comparing hydrocarbon-degrader MPNs with heterotrophic plate counts for several pure and mixed cultures. For some organisms, it seems likely that a single cell cannot initiate sufficient growth to produce a positive result. Thus, this and other hydrocarbon-degrader MPN procedures might underestimate the hydrocarbon-degrading population, even for culturable organisms.

A comparison of carbon/energy and complex nitrogen sources for bacterial sulphate-reduction: potential applications to bioprecipitation of toxic metals as sulphides.

Abstract: Detailed nutrient requirements were determined to maximise efficacy of a sulphate-reducing bacterial mixed culture for biotechnological removal of sulphate, acidity and toxic metals from waste waters. In batch culture, lactate produced the greatest biomass, while ethanol was more effective in stimulating sulphide production and acetate was less effective. The presence of additional bicarbonate and H2 only marginally stimulated sulphide production. The sulphide output per unit of biomass was greatest using ethanol as substrate. In continuous culture, ethanol and lactate were used directly as efficient substrates for sulphate reduction while acetate yielded only slow growth. Glucose was utilised following fermentation to organic acids and therefore had a deleterious effect on pH. Ethanol was selected as the most efficient substrate due to cost and efficient yield of sulphide. On ethanol, the presence of additional carbon sources had no effect on growth or sulphate reduction in batch culture but the presence of complex nitrogen sources (yeast extract or cornsteep) stimulated both. Cornsteep showed the strongest effect and was also preferred on cost grounds. In continuous culture, cornsteep significantly improved the yield of sulphate reduced per unit of ethanol consumed. These results suggest that the most efficient nutrient regime for bioremediation using sulphate-reducing bacteria required both ethanol as carbon source and cornsteep as a complex nitrogen source.

Microfungi from decaying leaves of two rain forest trees in Puerto Rico

Abstract: Fungal species richness and abundance were compared in leaf litter of two tree species,Guarea guidonia andManilkara bidentata, in the Luquillo Mountains of Puerto Rico. Four litter samples yielded a total of 3337 isolates, ranging from 591 to 1259 isolates/sample. The number of species/sample ranged from 134 to 228. Many uncommon litter hyphomycetes were recovered as well as coelomycetes, sterile strains, endophytes, and phytopathogens. Species-abundance distributions revealed a typical pattern of a few abundant species and a high proportion of rare species. Similarities in fungal species composition were not correlated with host species or with the site. Replicate samples examined by the moist chamber technique yielded a total of 24 species among the four litter samples. The particle filtration method indicated that leaves ofG. guidonia were more species-rich, while moist chambers indicated leaves ofM. bidentata were more species-rich. The moist chamber technique underestimated the number and species of viable fungi.

Continuous ethanol production by Zymomonas mobilis and Saccharomyces cerevisiae in biofilm reactors.

Abstract: Continuous ethanol fermentations were performed in duplicate for 60 days with Zymomonas mobilis ATCC 331821 or Saccharomyces cerevisiae ATCC 24859 in packed-bed reactors with polypropylene or plastic composite-supports. The plastic composite-supports used contained polypropylene (75%) with ground soybean-hulls (20%) and zein (5%) for Z. mobilis, or with ground soybean-hulls (20%) and soybean flour (5%) for S. cerevisiae. Maximum ethanol productivities of 536 g L-1 h-1 (39% yield) and 499 g L-1 h-1 (37% yield) were obtained with Z. mobilis on polypropylene and plastic composite-supports of soybean hull-zein, respectively. For Z. mobilis, an optimal yield of 50% was observed at a 1.92 h-1 dilution rate for soybean hull-zein plastic composite-supports with a productivity of 96 g L-1 h-1, whereas with polypropylene-supports the yield was 32% and the productivity was 60 g L-1 h-1. With a S. cerevisiae fermentation, the ethanol production was less, with a maximum productivity of 76 g L-1 h-1 on the plastic composite-support at a 2.88 h-1 dilution rate with a 45% yield. Polypropylene-support bioreactors were discontinued due to reactor plugging by the cell mass accumulation. Support shape (3-mm chips) was responsible for bioreactor plugging due to extensive biofilm development on the plastic composite-supports. With suspension-culture continuous fermentations in continuously-stirred benchtop fermentors, maximum productivities of 5 g L-1 h-1 were obtained with a yield of 24 and 26% with S. cerevisiae and Z. mobilis, respectively. Cell washout in suspension-culture continuous fermentations was observed at a 1.0 h-1 dilution rate. Therefore, for continuous ethanol fermentations, biofilm reactors out-performed suspension-culture reactors, with 15 to 100-fold higher productivities (g L-1 h-1) and with higher percentage yields for S. cerevisiae and Z. mobilis, respectively. Further research is needed with these novel supports to evaluate different support shapes and medium compositions that will permit medium flow, stimulate biofilm formation, reduce fermentation costs, and produce maximum yields and productivities.

Marine bacterial diversity as a resource for novel microbial products

Abstract: Marine bacteria are an important and relatively unexplored resource for novel microbial products. In this review, we discuss a number of issues relevant to the industrial potential of marine microorganisms including how marine and terrestrial bacteria differ, both physiologically and taxonomically, and what constitute reasonable expectations of the biosynthetic capabilities of marine bacteria relative to terrestrial bacteria and to marine macroorganisms. Also discussed is the concept that bacterial associations with marine plants and animals, which range from casual encounters to obligate symbioses, provide unique opportunities for bacterial adaptation. It is proposed that some of these adaptations would not be selected for in the absence of environmental parameters associated with the host, and that these adaptations can include the biosynthesis of unique metabolic products.

Production of high yields of docosahexaenoic acid by Thraustochytrium roseum ATCC 28210

Abstract: Culture conditions for growth and docosahexaenoic acid (DHA) production byThraustochytrium roseum ATCC 28210 were investigated with a view to increasing DHA titers. A medium was formulated (Medium 6) which produced a biomass and DHA content of 10.4 g L−1 and 1011 mg L−1, respectively, in a 5-day incubation. A fed-batch culture system was also developed which achieved biomass and DHA titers of 17.1 g L−1 and 2000 mg L−1, respectively, in 12 days.

Quest for wine yeasts—An old story revisited

Abstract: Numerous studies have described the yeast biota of grapes, and grape must in order to understand better the succession of yeasts during fermentation of wine. The origin of the wine yeasts has been rather controversial. By using more elaborate isolation methods, classical genetic analysis and electrophoretic karyotyping of monosporic clones, with this study, credible proof now exists that the vineyard is the primary source for the wine yeasts and that strains found on the grapes can be followed through the fermentation process.

Psychrophilic microorganisms and their cold-active enzymes

Abstract: The earth's vast and varied cold environments could be rich sources of psychrophilic microorganisms growing at 5°C or below. Unfortunately, the diversity, physiology and potential of these organisms have largely been overlooked. This article focuses on psychrophiles and their cold-active enzymes and emphasizes how future studies could give basic insight into protein structure and could yield industrially useful enzymes. It presents an overview of the characterization of psychrophiles and their growth properties, a summary of biochemical work with coldactive enzymes, a description of comparisons of enzymes with different temperature optima, and a preview of uses for cold-active enzymes in biotechnology.

Cyanobacteria carrying an smt-lux transcriptional fusion as biosensors for the detection of heavy metal cations

Abstract: The metal-responsivesmt operator/promoter region ofSynechococcus PCC7942 was fused to theluxCDABE genes ofVibrio fischeri. Plasmid DNA (pJLE23) carrying this fusion conferred metal ion-inducible luminescence to transformed cyanobacteria.Synechococcus PCC7942 (pJLE23) was sensitive to ZnCl2 concentrations within a range of 0.5–4 μM as demonstrated by induction of luminescence. Trace levels of CuSO4, and CdCl2 were also detected.

Molecular evidence for the presence of novel actinomycete lineages in a temperate forest soil

Abstract: PCR primers were designed to selectively recover partial (∼1100 bp) actinomycete 16S ribosomal DNA sequences from a temperate forest soil. A gene library was made and colony PCR was used to identify clones containing inserts. Unique clones were identified and partial or complete insert sequences were determined for 53 clones. Phylogenetic analyses revealed that 46 (87%) of the clones sampled contained 16S rDNA sequences which fell within the actinomycete radiation. The largest group of 34 sequences formed two closely related monophyletic groups in the 16S rRNA tree, which in turn formed a weakly supported sister group with the sequence fromActinomadura madurae. Four novel 16S rDNA lineages were detected inMycobacterium, one inPropionibacterium and one inCorynebacterium. Three novel sequences weakly grouped withSporichthya polymorpha. Two sequences formed an isolated lineage not closely related to any of the reference actinomycetes. Our results lend strong support to the hypothesis that cultured (and sequenced) actinomycetes do not adequately describe the diversity of this group in the environment.

Degradation of BTEX compounds in liquid media and in peat biofilters

Abstract: A mixed culture, enriched from Sphagnum peat moss, contaminated with gasoline vapours, degraded individual and mixed components of BTEX (benzene, toluene, ethylbenzene, xylene). Complete degradation of radiolabelled toluene by the mixed culture was observed in mineralisation studies. Individual isolates from a mixed culture containingPseudomonas maltophilia, P. testosteroni andP. putida biotype A exhibited contrasting BTEX degradation patterns. WhileP. putida biotype A degraded all of the BTEX compounds,P. maltophilia andP. testosteroni, appeared unable to degrade benzene and xylenes, respectively. When the peat, inoculated with the mixed culture, was used as a biofilter (6.2 cm diameter ×93 cm length) for degradation of toluene and ethylbenzene vapours, percentage removal efficiencies were 99 and 85, respectively. When the capacity of the biofilter to degrade a combination of BTEX compounds was evaluated, percentage removal efficiencies for toluene, ethylbenzene,p-xylene,o-xylene and benzene were 99, 85, 82, 80 and 78, respectively. The importance of using the mixed culture as an inoculum in the biofilter was established and also the relationship between contaminated vapour flow rate and percentage removal efficiency.