Anaerobic bioprocessing of organic wastes

Anaerobic digestion of dissolved, suspended and solid organics has rapidly evolved in the last decades but nevertheless still faces several scientific unknowns. In this review, some fundamentals of bacterial conversions and adhesion are addressed initially. It is argued in the light of G-values of reactions, and in view of the minimum energy quantum per mol, that anaerobic syntrophs must have special survival strategies in order to support their existence: redistributing the available energy between the partners, reduced end-product fermentation reactions and special cell-to-cell physiological interactions. In terms of kinetics, it appears that both reaction rates and residual substrate thresholds are strongly related to minimum G-values. These new fundamental insights open perspectives for efficient design and operation of anaerobic bioprocesses. Subsequently, an overview is given of the current anaerobic biotechnology. For treating wastewaters, a novel and high performance new system has been introduced during the last decade; the upflow anaerobic sludge blanket system (UASB). This reactor concept requires anaerobic consortia to grow in a dense and eco-physiologically well-organized way. The microbial principles of such granular sludge growth are presented. Using a thermodynamic approach, the formation of different types of aggregates is explained. The application of this bioprocess in worldwide wastewater treatment is indicated. Due to the long retention times of the active biomass, the UASB is also suitable for the development of bacterial consortia capable of degrading xenobiotics. Operating granular sludge reactors at high upflow velocities (5–6 m/h) in expanded granular sludge bed (EGSB) systems enlarges the application field to very low strength wastewaters (chemical oxygen demand < 1 g/l) and psychrophilic temperatures (10°C). For the treatment of organic suspensions, there is currently a tendency to evolve from the conventional mesophilic continuously stirred tank system to the thermophilic configuration, as the latter permits higher conversion rates and easier sanitation. Integration of ultrafiltration in anaerobic slurry digestion facilitates operation at higher volumetric loading rates and at shorter residence times. With respect to organic solids, the recent trend in society towards source separated collection of biowaste has opened a broad range of new application areas for solid state anaerobic fermentation.W. Verstraete and D. de Beer are with the Center for Environmental Sanitation, University of Gent, Coupure L 653, B-9000 Gent, Belgium; D. de Beer is also with the Max Plank Institut für Marine Mikrobiologie-Microzensor Group, Fahrenstrasse 1, 28359 Bremen, Germany. M. Pena is with the Groupo de Biotechnologia Ambiental, Departamento de Ingenieria Quimica, Universidad de Valladolid, Prado de la Magdalena, 47005 Valladolid, Spain. G. Lettinga is with the Department of Environmental Technology, Wageningen Agricultural University, Bomenweg 2, 6703 HD Wageningen, The Netherlands. P. Lens is with the Environmental Research Unit. Department of Microbiology, University College Galway, Galway, Ireland.     

合成生物制造2022

本文对2022年《生物工程学报》发表的与合成生物制造相关的综述和研究论文进行了评述,重点讨论了DNA测序、DNA合成、DNA编辑、基因表达调控和数学细胞模型等底层技术,酶的设计、改造和应用技术,化学品生物催化、氨基酸及其衍生物、有机酸、天然化合物、抗生素与活性肽、功能多糖、功能蛋白质等重要产品的生物制造技术,一碳化合物和生物质原料利用技术以及合成微生物组技术,以帮助读者从一个侧面了解合成生物制造相关技术和产业的发展情况。     

海洋氮循环中细菌的厌氧氨氧化

细菌厌氧氨氧化过程是在一类特殊细菌的厌氧氨氧化体内完成的以氨作为电子供体硝酸盐作为电子受体的一种新型脱氮反应.厌氧氨氧化菌的发现,改变人们对传统氮的生物地球化学循环的认识:反硝化细菌并不是大气中氮气产生的唯一生物类群.而且越来越多的证据表明,细菌厌氧氨氧化与全球的氮物质循环密切相关,估计海洋细菌的厌氧氨氧化过程占到全球海洋氮气产生的一半左右.由于氮与碳的循环密切相关,因此可以推测,细菌的厌氧氨氧化会影响大气中的二氧化碳浓度,从而对全球气候变化产生重要影响.另外,由于细菌厌氧氨氧化菌实现了氨氮的短程转化,缩短了氮素的转化过程,因此为开发更节约能源、更符合可持续发展要求的废水脱氮新技术提供了生物学基础.     

Novel autotrophic nitrogen removal system using gel entrapment technology

A pilot plant involving a nitritation-anammox process was operated for treating digester supernatant. In the preceding nitritation process, ammonium-oxidizing bacteria were immobilized in gel carriers, and the growth of nitrite-oxidizing bacteria was suppressed by heat-shock treatment. For the following anammox process, in order to maintain the anammox biomass in the reactor, a novel process using anammox bacteria entrapped in gel carriers was also developed. The nitritation performance was stable, and the average nitrogen loading and nitritation rates were 3.0 and 1.7 kg N m⁻³ d⁻¹, respectively. In the nitritation process, nitrate production was completely suppressed. For the anammox process, the startup time was about two months. Stable nitrogen removal was achieved, and an average nitrogen conversion rate of 5.0 kg N m⁻³ d⁻¹ was obtained. Since the anammox bacteria were entrapped in gel carriers, stable nitrogen removal performance was attained even at an influent suspended solids concentration of 1500 mg L⁻¹.     

Microbial degradation of the biocide polyhexamethylene biguanide: isolation and characterization of enrichment consortia and determination of degradation by measurement of stable isotope incorporation into DNA

AIMS: To isolate micro-organisms capable of utilizing polyhexamethylene biguanide (PHMB) as a sole source of nitrogen, and to demonstrate biodegradation of the biocide. METHODS AND RESULTS: Two consortia of bacteria were successfully enriched at the expense of PHMB, using sand from PHMB-treated swimming pools as inoculum. Both consortia were shown to contain bacteria belonging to the genera Sphingomonas, Azospirillum and Mesorhizobium. It was shown that the presence of both Sphingomonas and Azospirillum spp. was required for extensive growth of the consortia. In addition, the Sphingomonads were the only isolates capable of growth in axenic cultures dosed with PHMB. Using a stable isotope (15N)-labelled PHMB, metabolism of the biocide by both consortia was demonstrated. By comparing the level of 15N atom incorporation into bacterial DNA after growth on either 15N-PHMB or 15N-labelled NH4Cl, it was possible to estimate the percentage of PHMB biodegradation. CONCLUSIONS: The microbial metabolism of nitrogen from the biguanide moiety of PHMB has been demonstrated. It was revealed that Sphingomonas and Azospirillum spp. are the principal organisms responsible for growth at the expense of PHMB. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to demonstrate the microbial metabolism of PHMB.     

Effects of exogenously applied salicylic acid and putrescine alone and in combination with rhizobacteria on the phytoremediation of heavy metals and chickpea growth in sandy soil

The present attempt was made to study the role of exogenously applied salicylic acid (SA) and putrescine (Put) on the phytoremediation of heavy metals and on the growth parameters of chickpea grown in sandy soil. The SA and Put were applied alone as well as in combination with plant growth promoting rhizobacteria (PGPR). The PGPRs, isolated from the rhizosphere of chickpea, were characterized on the basis of colony morphology and biochemical traits through gram staining, catalase and oxidase tests, and identified by 16S-rRNA gene sequencing as Bacillus subtilis, Bacillus thuringiensis and Bacillus megaterium. The chickpea seeds were soaked in 24 h old fresh cultures of isolates for 2–3 h prior to sowing. The growth regulators (PGRs), SA and Put (150 mg/L), were applied to the seedlings as foliar spray at three-leaf stage. The result revealed that plants treated with SA and Put alone or in combination with PGPRs, significantly enhanced the accumulation of heavy metals in plant shoot. PGPR induces Ni accumulation in sensitive variety and Pb in both the varieties, the PGR in combination augment the bioremediation effects of PGPR and both sensitive and tolerant variety showed significant accumulation of Ni, Cd, and Pb. SA was more effective in accumulating Ni and Cd whereas, significant accumulation of Pb was recorded in Put. PGPRs further augmented the PGRs induced accumulation of heavy metals and macronutrients in chickpea shoot and in rhizosphere. SA increased the proline content of tolerant variety while decreasing the lipid peroxidation and proline content of the sensitive variety but decreased the stimulating effect of PGPR in proline production. Interactive effects of PGPR and PGRs are recommended for inducing phytoremediation in chickpea plants under drought stress.     

Design of mutualistic microbial consortia for stable conversion of carbon monoxide to value-added chemicals

Carbon monoxide (CO) is a promising carbon source for producing value-added biochemicals via microbial fermentation. However, its microbial conversion has been challenging because of difficulties in genetic engineering of CO-utilizing microorganisms and, more importantly, maintaining CO consumption which is negatively affected by the toxicity of CO and accumulated byproducts. To overcome these issues, we devised mutualistic microbial consortia, co-culturing Eubacterium limosum and genetically engineered Escherichia coli for the production of 3-hydroxypropionic acid (3-HP) and itaconic acid (ITA). During the co-culture, E. limosum assimilated CO and produced acetate, a toxic by-product, while E. coli utilized acetate as a sole carbon source. We found that this mutualistic interaction dramatically stabilized and improved CO consumption of E. limosum compared to monoculture. Consequently, the improved CO consumption allowed successful production of 3-HP and ITA from CO. This study is the first demonstration of value-added biochemical production from CO using a microbial consortium. Moreover, it suggests that synthetic mutualistic microbial consortium can serve as a powerful platform for the valorization of CO.     

Characterization of hydrocarbon-degrading bacteria isolated from oil-contaminated sediments in the Sultanate of Oman and evaluation of bioaugmentation and biostimulation approaches in microcosm experiments

Two oil-polluted sediments (PD and KH) were sampled from a coastal region in Oman for the isolation of hydrocarbon-degrading bacteria and for testing different bioremediation approaches. Fourty strains were isolated, eighteen were affiliated to Marinobacter whereas the rest belonged to Pseudomonas, Halomonas, Hahella and Alcanivorax. All strains grew well at 2–7% salinity and between 20 and 60 °C. The strains exhibited a better growth on long chain than on short chain alkanes. Biostimulation and bioaugmentation were compared in both sediments and oil biodegradation was followed by measuring CO₂ evolution and by gas chromatography (GC). The evolved CO₂ reached 0.45 ± 0.02 and 2.23 ± 0.07 mg CO₂ g⁻¹ sediment after 88 days in the untreated PD and KH sediments, respectively. While the addition of inorganic nutrients resulted in 1.2–3.7 fold increase in CO₂ evolution in both sediments, the addition of the bacterial consortium was only effective in the PD sediment. The maximum CO₂ evolution was measured when both nutrients and bacteria were added and this corresponded to a total oil mineralization of 2.6 ± 0.12 and 1.49 ± 0.04% of the initial oil after 88 days in the PD and KH sediments, respectively. GC analysis confirmed the CO₂ data and showed that most of the degraded compounds belonged to alkanes. We conclude that the Omani polluted sediments contain halotolerant and thermotolerant bacteria and biostimulation is more efficient than bioaugmentation for their cleanup.     

Efficient decolorization and detoxification of sulphonated azo dye Ponceau 4R by using single and mixed bacterial consortia

Abstract

The decolorization of toxic azo dye Ponceau 4R by three strains of bacteria Bacillus sp. strain AK1, Lysinibacillus sp. strain AK2 and Kerstersia sp. strain VKY1 individually and in consortia was studied. At optimal conditions, up to 95%, 93% and 87% of the dye was decolorized by the strains AK1, AK2 and VKY1, respectively, in 24?h at 200?mg/L of the dye. Decolorization of the dye was optimized for different parameters such as the concentration of dye, pH, temperature and NaCl concentration. These strains were able to decolorize Ponceau 4R up to an initial concentration of 800?mg/L in the pH range of 5–10, temperature 25–55?°C and NaCl concentration up to 30?g/L. The dye decolorization efficiency of these strains was further enhanced by using different consortia of AK1, AK2 and VKY1 in various combinations. The complete decolorization of the dye by a consortium was achieved within 18?h at 200?mg/L. The cell-free extract of these strains grown on this dye exhibited a remarkable activity of azoreductase which is involved in the breakage of the azo bond. The steady-state kinetics of azoreductase, validated the ping pong Bi-Bi mechanism of enzyme action. UV–Vis spectra, HPLC, FTIR and LC-MS analysis of the dye decolorized samples showed the formation of 4-aminonaphthalene-1-sulphonic acid and 5-amino-6-hydroxynaphthalene-2, 4-disulphonic acid as the products of azo bond breakage. The phytotoxicity test of decolorized sample revealed a considerable reduction in the toxicity in comparison with the parent dye.