Lipase-catalyzed selective synthesis of monolauroyl maltose using continuous stirred tank reactor

Monolauroyl maltose was synthesized by an immobilized lipase that catalyzed condensation of maltose and lauric acid in acetone using a batch reactor or a continuous stirred tank reactor. Mono- and di-lauroyl maltoses were identified by FT-IR, ¹H NMR, ¹³C NMR and MS. Monolauroyl maltose was selectively synthesized in a continuous stirred tank reactor and no diester was detected. The highest concentration of monolauroyl maltose at 28 mmol/l was obtained in 250 ml acetone when maltose was added at 4 g/d and the molar ratio of lauric acid to maltose was fixed at 4:1 at a flow rate of 0.15 ml/min for both influx and effluent without supplement of fresh molecular sieve.     

Anaerobic methanethiol degradation and methanogenic community analysis in an alkaline (pH 10) biological process for liquefied petroleum gas desulfurization

Anaerobic methanethiol (MT) degradation by mesophilic (30 degrees C) alkaliphilic (pH 10) communities was studied in a lab-scale Upflow Anaerobic Sludge Bed (UASB) reactor inoculated with a mixture of sediments from the Wadden Sea (The Netherlands), Soap Lake (Central Washington), and Russian soda lakes. MT degradation started after 32 days of incubation. During the first 252 days, complete degradation was achieved till a volumetric loading rate of 7.5 mmol MT/L/day, and sulfide, methane, and carbon dioxide were the main reaction products. Temporary inhibition of MT degradation occurred after MT peak loads and in the presence of dimethyl disulfide (DMDS), which is the autooxidation product of MT. From day 252 onwards, methanol was dosed to the reactor as co-substrate at a loading rate of 3-6 mmol/L/day to stimulate growth of methylotrophic methanogens. Methanol was completely degraded and also a complete MT degradation was achieved till a volumetric loading rate of 13 mmol MT/L/day (0.77 mmol MT/gVSS/day). However, from day 354 till the end of the experimental run (day 365), acetate was formed and MT was not completely degraded anymore, indicating that methanol-degrading homoacetogenic bacteria had partially outcompeted the methanogenic MT-degrading archea. The archeal community in the reactor sludge was analyzed by DGGE and sequencing of 16S rRNA genes. The methanogenic archea responsible for the degradation of MT in the reactor were related to Methanolobus oregonensis. A pure culture, named strain SODA, was obtained by serial dilutions in medium containing both trimethyl amine and dimethyl sulfide (DMS). Strain SODA degraded MT, DMS, trimethyl amine, and methanol. Flow sheet simulations revealed that for sufficient MT removal from liquefied petroleum gas, the extraction and biological degradation process should be operated above pH 9.     

Pretreatment of textile dyeing wastewater using an anoxic baffled reactor

A study on pretreatment of textile dyeing wastewater was carried out using an anoxic baffled reactor (ABR) at wastewater temperatures of 5-31.1 degrees C. When hydraulic retention time (HRT) was 8h, the color of outflow of ABR was only 40 times at 5 degrees C and it could satisfy the professional discharge standard (grade-1) of textile and dyeing industry of China (GB4287-92). The total COD removal efficiency of ABR was 34.6%, 47.5%, 50.0%, 53.3%, 54.7% and 58.1% at 5, 9.7, 14.9, 19.7, 23.5 and 31.1 degrees C, respectively. Besides, after the wastewater being pre-treated by ABR when HRT was 6h and 8h, the BOD5/COD value rose from 0.30 of inflow to 0.46 of outflow and from 0.30 of inflow to 0.40 of outflow, respectively. Experimental results indicated that ABR was a very feasible process to decolorize and pre-treat the textile dyeing wastewater at ambient temperature. Moreover, a kinetic simulation of organic matter degradation in ABR at six different wastewater temperatures was carried through. The kinetic analysis showed the organic matter degradation was a first-order reaction. The reaction activation energy was 19.593 kJ mol(-1) and the temperature coefficient at 5-31.1 degrees C was 1.028.     

填充床反应器中酶法连续合成甘油二酯的研究

近年来,1,3-甘油二酯(DAG)由于其广泛用途及健康作用日益受到人们的重视。报道了一种无溶剂条件下填充床反应器中连续酶促合成1,3-DAG的方法。研究了填充柱的长径比、进料体积流速、温度、底物摩尔比对酯化率和1,3-DAG产量的影响。结果表明固定化酶填充柱长径比7.8,亚油酸、甘油摩尔比1∶2 ,进料速度1.2mL/min ,65℃条件下酯化反应可实现脂肪酸酯化率、1,3-DAG纯度及生产效率的统一。填充床反应器中固定化酶连续催化酯化反应的一个主要问题即体系水分清除困难。实验研究了采用过量甘油吸附脱水的可行性,亚油酸、甘油摩尔比为1∶2时,可明显改善固定化酶的稳定性,增加LipozymeRMIM的使用寿命。连续运行10d ,残余酶活仍保持在80 %以上,而对照组则仅为52%。     

Anaerobic Digestion of Dairy Manure in a Hybrid Reactor with Biogas Recirculation

Summary A novel anaerobic hybrid reactor (AHR) configuration incorporating floating support media for biomass immobilization and biogas recirculation for enhanced mixing was used for anaerobic digestion of dairy manure. No pretreatment or solid liquid separation was applied. The reactor was operated at high influent volatile solids (VS) and organic loading rates (OLR) of up to 9.87% and 7.30 g VS/l day, respectively. After 149 days of continuous operation the results revealed that a high amount (38.1 g VSS) of biomass was able to attach itself to the support medium being used. The investigated AHR configuration achieved COD, BOD, TS, and VS removal efficiencies of 48–63, 64–78, 55–65, and 59–68%, respectively, at a hydraulic retention time (HRT) of 15 days. The corresponding average methane production value obtained in this study was 0.191 l/g VS added.     

A study in UF-membrane reactor on activity and stability of nitrile hydratase from Microbacterium imperiale CBS 498-74 resting cells for propionamide production

The bioconversion of propionitrile to propionamide was catalysed by nitrile hydratase (NHase) using resting cells of Microbacterium imperiale CBS 498-74 (formerly, Brevibacterium imperiale). This microorganism, cultivated in a shake flask, at 28 °C, presented a specific NHase activity of 34.4 U mg_DCW⁻¹ (dry cell weight). The kinetic parameters, K_m and V_max, tested in 50 mM sodium phosphate buffer, pH 7.0, in the propionitrile bioconversion was evaluated in batch reactor at 10 °C and resulted 21.6 mM and 11.04 μmol min⁻¹ mg_DCW⁻¹, respectively. The measured apparent activation energy, 25.54 kJ mol⁻¹, indicated a partial control by mass transport, more likely through the cell wall.

UF-membrane reactors were used for kinetic characterisation of the NHase catalysed reaction. The time dependence of enzyme deactivation on reaction temperature (from 5 to 25 °C), on substrate concentrations (from 100 to 800 mM), and on resting cell loading (from 1.5 to 200 μg  ml⁻¹) indicated: lower diffusional control (E_a=37.73 kJ mol⁻¹); and NHase irreversible damage caused by high substrate concentration. Finally, it is noteworthy that in an integral reactor continuously operating for 30 h, at 10 °C, 100% conversion of propionitrile (200 mM) was attained using 200 μg  ml⁻¹ of resting cells, with a maximum volumetric productivity of 0.5 g l⁻¹ h⁻¹.     

Protein refolding in an oscillatory flow reactor

We demonstrate that an oscillatory flow reactor is a viable reactor for protein refolding via direct dilution. The mixing characteristics of the oscillatory flow reactor are well described and controllable and, importantly, can be scaled-up to process scale without a loss of mixing efficiency. This makes the oscillatory flow reactor an attractive alternative to conventional stirred-tank reactors for process-scale renaturation.     

Biotransformation of glucose to gluconic acid by Aspergillus niger—study of mass transfer in an airlift bioreactor

A glucose–gluconic acid biotransformation system was suggested for the experimental study of oxygen transfer in bioreactors. This biosystem was used for the investigation of the effect of the flow rate and biomass concentration on the volumetric oxygen transfer coefficient k_La in a 10 dm³ internal-loop airlift bioreactor. For this purpose, the fermentation broth of the mycelial strain Aspergillus niger was employed, representing a three-phase system, where bubbles come into contact with dense rigid pellets. The results showed that the presented biotransformation system can be successfully utilised for the determination of the oxygen transfer rate in airlift bioreactors. The experiments showed a strong positive influence of the air flow rate on the rate (r_Glu), specific rate of gluconic acid production (k_Glu/X) as well as on the volumetric oxygen transfer coefficient (k_La). This confirmed an expected limitation of production rate by the oxygen transport from the gas to the liquid phase in the whole range of air flow rates applied. Moreover, consistent curves of the production rate r_Glu and k_La values vs. biomass concentration c_X (amount of enzymes) were observed. These exhibited a local maximum for c_X equal to 6.68 g dm⁻³. On the other hand, the specific production rate monotonously decreased with increasing biomass concentration. A decline of k_La values at higher c_X values was attributed to a bubble coalescence promoting effect of mycelial pellets.     

Hydrodynamics and mass transfer in a tubular airlift photobioreactor

In photobioreactors, which are usually operated under light limitation,sufficient dissolved inorganic carbon must be provided to avoid carbonlimitation. Efficient mass transfer of CO₂ into the culture mediumisdesirable since undissolved CO₂ is lost by outgassing. Mass transferof O₂ out of the system is also an important consideration, due tothe need to remove photosynthetically-derived O₂ before it reachesinhibitory concentrations. Hydrodynamics (mixing characteristics) are afunctionof reactor geometry and operating conditions (e.g. gas and liquid flow rates),and are a principal determinant of the light regime experienced by the culture.This in turn affects photosynthetic efficiency, productivity, and cellcomposition. This paper describes the mass transfer and hydrodynamics within anear-horizontal tubular photobioreactor. The volume, shape and velocity ofbubbles, gas hold-up, liquid velocity, slip velocity, axial dispersion,Reynoldsnumber, mixing time, and mass transfer coefficients were determined intapwater,seawater, and algal culture medium. Gas hold-up values resembled those ofvertical bubble columns, and the hydraulic regime could be characterized asplug-flow with medium dispersion. The maximum oxygen mass transfer coefficientis approximately 7 h^–1. A regime analysisindicated that there are mass transfer limitations in this type ofphotobioreactor. A methodology is described to determine the mass transfercoefficients for O₂ stripping and CO₂ dissolution whichwould be required to achieve a desired biomass productivity. This procedure canassist in determining design modifications to achieve the desired mass transfercoefficient.     

Overall assessment of Monodus subterraneus cultivation and EPA production in outdoor helical and bubble column reactors

The outdoor production of Monodus subterraneus wasstudied in bubble column and helical reactors, mainly analysing the influenceofdilution rate, air flow rate and solar irradiance on growth rate andbiochemicalcomposition. Photoinhibition and photo-oxidation phenomena were also analysed.The cultures were stressed at high solar irradiance and dissolved oxygenconcentrations. A clear relationship between stress of the cultures and thefluorescence from PSII measurements was observed, the Fv/Fm ratio being lowerinthe helical reactor than in the bubble column. Growth rate and biomassproductivity were both a function of the average irradiance and the Fv/Fmratio;maximum values of 0.040 h^–1 and 0.54 gL^–1 d^–1 were measured. The influenceofphotoinhibition and average irradiance was modelled, the model also fitting theexperimental data reported by another author. The chlorophyll contenthyperbolically decreased, whereas the carotenoid content decreased linearlywiththe average irradiance. The higher the dilution rate the higher the protein andcarbohydrate content of the biomass, and the lower the lipid content. Theeicosapentaenoic acid (EPA) content ranged from 2.3 to 3.2% d.wt, the higherthe dilution rate, the lower EPA content, although the higher the EPAproportion. Maximum EPA productivity was only 9 mg L^–1d^–1, due to the stress to which the cultures wereexposed.