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Protein extraction from tea leaf residues at high yield
New Biotechnology · Volume 29S · September 2012
References 1. Hetényi. J Chem Technol Biotechnol 2010;85:872–7. 2. Hetényi. Chem Eng Proc: Proc Intens 2011;50:293–9. 3. Hetényi, K. (2011) Technological improvements of a biorefinery. PhD thesis, Budapest BME.
http://dx.doi.org/10.1016/j.nbt.2012.08.010 Oral 1.2.04 Direvo BluCon-P: production of biofuels and -chemicals from pretreated grass and wood in a one-step consolidated bioprocess Albrecht Läufer ∗ , Vitaly Svetlichnyi, Simon Curvers Direvo Industrial Biotechnology GmbH, Nattermannallee 1, D-50829 Cologne, Germany Direvo’s consolidated bioprocess BluCon-P targets lowest possible manufacturing cost by consolidating all relevant process steps for conversion of lignocellulosic feedstock into biofuel and biochemicals into one unit operation. It was designed to • Be feedstock agnostic • Allow easy separation of product • Avoid secondary infections. Several hundreds of samples from individual microbial ecosystems were screened and a versatile set of new, extremely thermophilic microbes has been found. Based on these new strains substrate- and product-tailored production processes are being designed. The following process steps are integrated into one unit operation: • Production of hemi/cellulolytic enzymes • Hydrolysis of celluloses and hemicelluloses • Conversion of C6 sugars and C5 sugars to products: ethanol, lactic acid, acetic acid • In situ ethanol capture Product concentrations of 0.3% vol. for ethanol, and 2.5 g/L for lactic acid could be achieved under currently optimal conditions using the wild type strains. A broad range of different, pretreated substrates including perennial grasses, agricultural residues as well as hard- and softwood was converted with high substrate yields of up to 96% in the case of steam pretreated miscanthus. http://dx.doi.org/10.1016/j.nbt.2012.08.011 Oral 1.2.05 Protein extraction from tea leaf residues at high yield Chen Zhang ∗ , Marieke E. Bruins, Johan P.M. Sanders Biobased Commodity Chemicals Group, Bornse Weilanden 9, 6708WG Wageningen, Wageningen UR, The Netherlands Leaf protein (LP) is considered to be a potential resource for feed or food since 1960s, but its applications are limited due to high proportion of insoluble protein and low cost-efficient processing. To overcome these problems, the alkaline extraction method and correlated parameters are re-evaluated by using green tea leaf residue (GTR) as starting material. In this study, protein extraction
of GTR could be maximized up to 95%. During extraction, the protein extractability to GTR extractability ratio was constant at either 0.28 or 0.57, which indicates two different extraction mechanisms. In the extraction, temperature and NaOH are crucial and interacting parameters that affect extraction efficiency and final protein molecular weight. By controlling the process with these two parameters it is possible to get high recovery of proteins in the extracts that are still at high molecular weight, which benefits the subsequent recovery. The economical optimal one-step method produces 53% pure protein from tea leaves at only 130D/ton product. As the amino acid composition of tea protein extracts are comparable to that of soybean meal, the price of tea leaf protein can be up to 400D/ton. In addition, this new technology can be also applied to other leaves, which increases the chances for cost-efficient applications of leaf protein. http://dx.doi.org/10.1016/j.nbt.2012.08.012 Stream: White – Industrial Biotechnology, Session: Biobased Chemicals & Materials Oral 1.3.01 Industrial biotechnology: directing innovation in chemistry Bernhard Hauer Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany Biotechnology promised to be a major driver to the challenging problems we face in the 21st century. Under the term of industrial or white biotechnology significant contributions are expected for the quest of new system solutions in chemistry. This intention should implement novel as well as drop in chemicals and materials based on a renewable raw material basis. However we have to expand our portfolio of biocatalyst towards new reactions to boost the impact of biotechnology [1]. We postulate that the pool of biocatalytic reactions can be greatly expanded by harnessing the catalytic machinery of known enzymes. I will present a few selected examples from our studies with monooxygenases [2], enoate reductases [3] and cyclases [4]. Squalene hopene cyclases (SHCs), for example, catalyse the Brønsted acid initiated cyclisation of squalene to hopene via cationic intermediates. Bearing in mind the large diversity of reactions catalysed by Brønsted acid catalysts in synthetic organic chemistry these enzymes should be able not only to convert non-natural substrates (substrate promiscuity) but also catalyse non-natural reactions (catalytic promiscuity). Employing this approach we show that cyclases can catalyse Friedel-Crafts alkylations and the synthesis of different heterocyclic compounds. Our results indicate that the catalytic machinery of SHCs can be exploited for general Brønsted acid catalysis in a chiral environment [5]. The next step will be the combination of novel enzymes to entire new biosynthetic pathways to enable us to manufacture ‘new-to-nature’ products. With this perspective, we can anticipate the relevance of biotechnology to chemistry to accelerate even further. www.elsevier.com/locate/nbt S7
References 1. Hetényi. J Chem Technol Biotechnol 2010;85:872–7. 2. Hetényi. Chem Eng Proc: Proc Intens 2011;50:293–9. 3. Hetényi, K. (2011) Technological improvements of a biorefinery. PhD thesis, Budapest BME.
http://dx.doi.org/10.1016/j.nbt.2012.08.010 Oral 1.2.04 Direvo BluCon-P: production of biofuels and -chemicals from pretreated grass and wood in a one-step consolidated bioprocess Albrecht Läufer ∗ , Vitaly Svetlichnyi, Simon Curvers Direvo Industrial Biotechnology GmbH, Nattermannallee 1, D-50829 Cologne, Germany Direvo’s consolidated bioprocess BluCon-P targets lowest possible manufacturing cost by consolidating all relevant process steps for conversion of lignocellulosic feedstock into biofuel and biochemicals into one unit operation. It was designed to • Be feedstock agnostic • Allow easy separation of product • Avoid secondary infections. Several hundreds of samples from individual microbial ecosystems were screened and a versatile set of new, extremely thermophilic microbes has been found. Based on these new strains substrate- and product-tailored production processes are being designed. The following process steps are integrated into one unit operation: • Production of hemi/cellulolytic enzymes • Hydrolysis of celluloses and hemicelluloses • Conversion of C6 sugars and C5 sugars to products: ethanol, lactic acid, acetic acid • In situ ethanol capture Product concentrations of 0.3% vol. for ethanol, and 2.5 g/L for lactic acid could be achieved under currently optimal conditions using the wild type strains. A broad range of different, pretreated substrates including perennial grasses, agricultural residues as well as hard- and softwood was converted with high substrate yields of up to 96% in the case of steam pretreated miscanthus. http://dx.doi.org/10.1016/j.nbt.2012.08.011 Oral 1.2.05 Protein extraction from tea leaf residues at high yield Chen Zhang ∗ , Marieke E. Bruins, Johan P.M. Sanders Biobased Commodity Chemicals Group, Bornse Weilanden 9, 6708WG Wageningen, Wageningen UR, The Netherlands Leaf protein (LP) is considered to be a potential resource for feed or food since 1960s, but its applications are limited due to high proportion of insoluble protein and low cost-efficient processing. To overcome these problems, the alkaline extraction method and correlated parameters are re-evaluated by using green tea leaf residue (GTR) as starting material. In this study, protein extraction
of GTR could be maximized up to 95%. During extraction, the protein extractability to GTR extractability ratio was constant at either 0.28 or 0.57, which indicates two different extraction mechanisms. In the extraction, temperature and NaOH are crucial and interacting parameters that affect extraction efficiency and final protein molecular weight. By controlling the process with these two parameters it is possible to get high recovery of proteins in the extracts that are still at high molecular weight, which benefits the subsequent recovery. The economical optimal one-step method produces 53% pure protein from tea leaves at only 130D/ton product. As the amino acid composition of tea protein extracts are comparable to that of soybean meal, the price of tea leaf protein can be up to 400D/ton. In addition, this new technology can be also applied to other leaves, which increases the chances for cost-efficient applications of leaf protein. http://dx.doi.org/10.1016/j.nbt.2012.08.012 Stream: White – Industrial Biotechnology, Session: Biobased Chemicals & Materials Oral 1.3.01 Industrial biotechnology: directing innovation in chemistry Bernhard Hauer Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany Biotechnology promised to be a major driver to the challenging problems we face in the 21st century. Under the term of industrial or white biotechnology significant contributions are expected for the quest of new system solutions in chemistry. This intention should implement novel as well as drop in chemicals and materials based on a renewable raw material basis. However we have to expand our portfolio of biocatalyst towards new reactions to boost the impact of biotechnology [1]. We postulate that the pool of biocatalytic reactions can be greatly expanded by harnessing the catalytic machinery of known enzymes. I will present a few selected examples from our studies with monooxygenases [2], enoate reductases [3] and cyclases [4]. Squalene hopene cyclases (SHCs), for example, catalyse the Brønsted acid initiated cyclisation of squalene to hopene via cationic intermediates. Bearing in mind the large diversity of reactions catalysed by Brønsted acid catalysts in synthetic organic chemistry these enzymes should be able not only to convert non-natural substrates (substrate promiscuity) but also catalyse non-natural reactions (catalytic promiscuity). Employing this approach we show that cyclases can catalyse Friedel-Crafts alkylations and the synthesis of different heterocyclic compounds. Our results indicate that the catalytic machinery of SHCs can be exploited for general Brønsted acid catalysis in a chiral environment [5]. The next step will be the combination of novel enzymes to entire new biosynthetic pathways to enable us to manufacture ‘new-to-nature’ products. With this perspective, we can anticipate the relevance of biotechnology to chemistry to accelerate even further. www.elsevier.com/locate/nbt S7