Thermophilic hydrogen production from cellulose with rumen fluid enrichment cultures: Effects of different heat treatments
Elevated temperatures (52, 60 and 65 °C) were used to enrich hydrogen producers on cellulose from cow rumen fluid. Methanogens were inhibited with two different heat treatments. Hydrogen production was considerable at 60 °C with the highest H2 yield of 0.44 mol-H2 mol-hexose -1 (1.93 mol-H2 mol-hexose-degraded-1) as obtained without heat treatment and with acetate and ethanol as the main fermentation products. H2 production rates and yields were controlled by cellulose degradation that was at the highest 21%. The optimum temperature and pH for H2 production of the rumen fluid enrichment culture were 62 °C and 7.3, respectively. The enrichments at 52 and 60 °C contained mainly bacteria from …
Production and characterization of the recombinant Sphingomonas chlorophenolica pentachlorophenol 4-monooxygenase.
Abstract Pentachlorophenol 4-monooxygenase (PCP4MO) from Sphingomonas chlorophenolica is a flavoprotein that hydroxylates PCP in the presence of NADPH and oxygen. In order to investigate the structure and function of active site, recombinant PCP4MO (rePCP4MO) was produced in Escherichia coli as a glutathione S-transferase (GST) fusion protein. Moreover, a tobacco etch virus (TEV) protease cleavage site (EKLYFQG) was introduced into GST-PCP4MO and a his-tagged TEV protease was employed. Hence, a two-step purification protocol was developed which allowed obtaining 15–20 mg of rePCP4MO from 1 L culture. The rePCP4MO revealed identity with native enzyme by SDS–PAGE and N-terminal sequence analy…
Diversity of chlorophenol-degrading bacteria isolated from contaminated boreal groundwater
Chlorophenol-degrading bacteria from a long-term polluted groundwater aquifer were characterized. All isolates degraded 2,4,6-trichlorophenol and 2,3,4,6-tetrachlorophenol at concentrations detected in the contaminated groundwater (10 mg 1(-1)). Pentachlorophenol was degraded by three isolates when present alone. In two gram-positive isolates, 2,3,4,6-tetrachlorophenol was required as an inducer for the degradation of pentachlorophenol. The gram-positive isolates were sensitive to pentachlorophenol, with an IC50 value of 5 mg/l. Isolates belonging to the Cytophaga/Flexibacter/Bacteroides phylum had IC50 values of 25 and 63 mg/l. Isolates belonging to alpha-, beta- and gamma-Proteobacteria g…
Effects of heat treatment on hydrogen production potential and microbial community of thermophilic compost enrichment cultures
Cellulosic plant and waste materials are potential resources for fermentative hydrogen production. In this study, hydrogen producing, cellulolytic cultures were enriched from compost material at 52, 60 and 70°C. Highest cellulose degradation and highest H(2) yield were 57% and 1.4 mol-H(2) mol-hexose(-1) (2.4 mol-H(2) mol-hexose-degraded(-1)), respectively, obtained at 52°C with the heat-treated (80°C for 20 min) enrichment culture. Heat-treatments as well as the sequential enrichments decreased the diversity of microbial communities. The enrichments contained mainly bacteria from families Thermoanaerobacteriaceae and Clostridiaceae, from which a bacterium closely related to Thermoanaerobiu…
Isolation and Characterization of Novosphingobium sp. Strain MT1, a Dominant Polychlorophenol-Degrading Strain in a Groundwater Bioremediation System
ABSTRACT A high-rate fluidized-bed bioreactor has been treating polychlorophenol-contaminated groundwater in southern Finland at 5 to 8°C for over 6 years. We examined the microbial diversity of the bioreactor using three 16S ribosomal DNA (rDNA)-based methods: denaturing gradient gel electrophoresis, length heterogeneity-PCR analysis, and restriction fragment length polymorphism analysis. The molecular study revealed that the process was dependent on a stable bacterial community with low species diversity. The dominant organism, Novosphingobium sp. strain MT1, was isolated and characterized. Novosphingobium sp. strain MT1 degraded the main contaminants of the groundwater, 2,4,6-trichloroph…
Spatial and temporal changes in Actinobacterial dominance in experimental artificial groundwater recharge.
Abstract Artificial groundwater recharge (AGR) is used in the drinking water industry to supplement groundwater resources and to minimise the use of chemicals in water treatment. This study analysed the spatial and temporal changes of microbial communities in AGR using two test systems: a nutrient-amended fluidized-bed reactor (FBR) and a sand column. Structural changes in the feed lake water (Lake Roine), FBR, and sand column bacterial communities were determined by denaturing gradient gel electrophoresis (DGGE) and the length heterogeneity analysis of amplified 16S rRNA genes (LH-PCR). Two clone libraries were created to link the LH-PCR results to the dominant bacterial groups. The lake w…
Impact of crop species on bacterial community structure during anaerobic co-digestion of crops and cow manure
The bacterial communities in three continuously stirred tank reactors co-digesting cow manure with grass silage, oat straw, and sugar beet tops, respectively, were investigated by 16S rRNA gene-based fingerprints and clone libraries. The analyses revealed both clearly distinct and similar phylotypes in the bacterial communities between the reactors. The major groups represented in the three reactors were Clostridia, unclassified Bacteria, and Bacteroidetes. Phylotypes affiliated with Bacilli or Deltaproteobacteria were unique to the sugar beet and straw reactor, respectively. Unclassified Bacteria dominated in sugar beet reactor while in the straw and grass reactor Clostridia was the domina…
Microbial Adaptation to Boreal Saturated Subsurface: Implications in Bioremediation of Polychlorophenols
Saturated subsurface environments pose challenges to the intrinsic microbiology. Prevailing environmental conditions (temperature, pH, bioavailability of substrates and nutrients) affect microbial biodegradation activity, which is often favored by certain redox conditions. Microbial adaptation in each redox environment proceeds by selection and enrichment of indigenous bacteria, evolution of novel catabolic pathways and horizontal gene transfer (Wilson et al. 1985; van der Meer et al. 1998; Tiirola et al. 2002b). Formation of biofilms enables microbial retainment, co-operation among microorganisms and enhanced gene transfer among organisms (Singh et al. 2006). Chlorophenols are toxic and pe…