On-Line Monitoring of Enzymatic Degumming of Soybean Oil Using Near-Infrared Spectroscopy.

Degumming is an oil refinement process in which the naturally occurring phospholipids in crude vegetable oils are removed. Enzymatic degumming results in higher oil yield and more cost-efficient processing compared to traditional degumming processes using only water or acid. Phospholipase C hydrolyses phospholipids into diglycerides and phosphate groups during degumming. The diglyceride content can therefore be considered a good indicator of the state of the enzymatic reaction. This study investigates the use of near-infrared (NIR) spectroscopy and chemometrics to monitor the degumming process by quantifying diglycerides in soybean oil in both off-line and on-line settings. Fifteen enzymatic degumming lab scale batches originating from a definitive screening design (with varying water, acid, and enzyme dosages) were investigated with the aim to develop a NIR spectroscopy prediction method. By applying tailored preprocessing and variable selection methods, the diglyceride content can be predicted with a root mean square error of prediction of 0.06% (w/w) for the off-line set-up and 0.07% (w/w) for the on-line set-up. The results show that the diglyceride content is a good indicator of the enzyme performance and that NIR spectroscopy is a suitable analytical technique for robust real-time diglyceride quantification.

Removal of hard COD from acidic eucalyptus kraft pulp bleach plant effluent streams using oxidoreductases.

The bleach plant of a pulp and paper (P&P) mill presents a major source of wastewater containing toxic organic matter characterized as chemical oxygen demand (COD). Due to their high oxidizing power, oxidoreductases hold promise to be a key solution for the removal of dissolved organic material. Here, four oxidoreductases from different enzyme families were selected to treat bleach plant effluents. Haloperoxidase treatment of the final effluent resulted in the highest levels of decolorization (71%) and reduction of aromatic compounds (36%). Using single compound analysis, 27 low molecular weight compounds were found to be persistent throughout the wastewater treatment process and, therefore, classified as hard COD. The tested enzymes efficiently removed several of the identified COD compounds. Hence, this study suggests that the application of oxidoreductases will serve as an environmental-friendly solution for reducing waste from P&P production.

Enhanced efficiency of industrial-scale anaerobic digestion by the addition of glycosidic enzymes.

The results of the action of two glycosidic enzymes added to an anaerobic digester processing mixed wastewater sludge are reported in this paper. The experiment was performed at a continuously operating full-scale wastewater treatment plant during a 6-month period. The addition of the enzyme mixture resulted in improved gas production and dewatering properties. Dewatering experiments, both on enzyme-treated and reference sludge on the local centrifuges, indicated that the carbonaceous matter was decreased by 7% in enzyme-treated sludge, and the average dry solids increased from 27% (reference) to 31% in enzyme-treated sludge. The polymer dosage was decreased from 8 to 5 kg/t dry solids. Additionally, practical experiences were gathered concerning the transformation from static laboratory-scale batch experiments to a dynamic continuous industrial scale setup. Preliminary cost calculations showed beneficial aspects and economical feasibility of enzyme addition to an anaerobic sludge digestion process.

The function of cation-binding agents in the enzymatic treatment of municipal sludge.

Treatment of sludge with enzymes has previously been shown to efficiently release organic matter. However, the added enzymes were partially adsorbed to, entrapped by or bound to the sludge structure. Simultaneous decrease of enzymes activities was observed. Reduced adsorption and more effective, lower, enzyme dose was achieved in sludge pre-treated with three cation-binding agents. The enzymatic solubilisation of sludge was improved by 150%, 240% and 290%, by 50mM sodium tripolyphosphate (STPP), 25mM citric acid (CA) or 50mM ethylenediaminetetraacetate (EDTA), respectively. With cation binders, the lower relative enzyme dose 0.2 (13.7mg/g total solids (TS)) released 3.5 times higher COD than enzyme dose 1 (68.5mg/g TS) alone. In the presence of 25mM CA, 75% added protease remained soluble. In the presence of 50mM CA, EDTA or STPP, 50% of alpha-amylase and cellulase remained soluble. At 200mM STPP, alpha-amylase was inactive, and the efficiency of enzymatic sludge hydrolysis decreased. CA was the most effective of the three cation-binding agents tested. It is biodegradable and can be produced endogenously by the microorganisms in sludge. CA has the greatest potential for the practical application to enhance biogas production. This paper reports on the possible mechanisms of enzymes adsorption to the sludge matrix and possible methods of decreasing the adsorption. We suggest that steric hindrances were responsible for the decreased enzymatic sludge solubilisation and that polyvalent metal ions were directly involved in adsorption of enzymes to sludge matrix. The addition of cation binders eliminated both phenomena and thereby improved the enzymatic solubilisation of sludge.

Poly(3-hydroxybutyrate) biosynthesis in the biofilm of Alcaligenes eutrophus, using glucose enzymatically released from pulp fiber sludge.

Glucose, enzymatically released from pulp fiber sludge, was combined with inorganic salts and used as a growth medium for Alcaligenes eutrophus, a gram-negative strain producing poly(3-hydroxybutyrate) (PHB). By controlling the concentrations of the inorganic salts in the growth medium, almost 78% of the cell mass was converted to pure PHB. Efforts were made to find conditions for bacterial growth in the form of a biofilm on a cheap and reusable carrier. A number of positively charged carriers were tested, and the anion exchanger DEAE-Sephadex A-25 was chosen as a microcarrier for packed-bed biofilm cultures of A. eutrophus. Conditions for attachment, growth, and detachment were established. Biofilm formation on the microcarrier is strongly dependent on the ionic strength of the attachment medium. In order to achieve formation of the biofilm and its recovery from the microcarrier, the ionic strengths of the attachment and the detachment media were varied. Low ionic strength was tested for attachment, and high ionic strength was tested for detachment. Although biofilm formation in the packed-bed reactor is limited, the volumetric yield of cells based on the void volume of the packed bed is comparable with the batch culture yield.