Plant cell walls to ethanol

Conversion of plant cell walls to ethanol constitutes second generation bioethanol production. The process consists of several steps: biomass selection/genetic modification, physiochemical pretreatment, enzymatic saccharification, fermentation and separation. Ultimately, it is desirable to combine as many of the biochemical steps as possible in a single organism to achieve CBP (consolidated bioprocessing). A commercially ready CBP organism is currently unreported. Production of second generation bioethanol is hindered by economics, particularly in the cost of pretreatment (including waste management and solvent recovery), the cost of saccharification enzymes (particularly exocellulases and endocellulases displaying kcat ~1?s-1 on crystalline cellulose), and the inefficiency of co-fermentation of 5- and 6-carbon monosaccharides (owing in part to redox cofactor imbalances in Saccharomyces cerevisiae).     

Plant biotechnology

Book Review

Plant biotechnologyS.-D. Kung and C.J. Arntzen (Eds.) Boston: Butterworths, 1989. xxi + 423 pages. £60.00. ISBN 0-409-90068-0     

Plant biotechnology for crop improvement

The typical crop improvement cycle takes 10–15 years to complete and includes germplasm manipulations, genotype selection and stabilization, variety testing, variety increase, proprietary protection and crop production stages. Plant tissue culture and genetic engineering procedures that form the basis of plant biotechnology can contribute to most of these crop improvement stages. This review provides an overview of the opportunities presented by the integration of plant biotechnology into plant improvement efforts and raises some of the societal issues that need to be considered in their application.     

Plant cell walls: wall-associated kinases and cell expansion

Arabidopsis has a family of five cell wall-associated protein kinases (WAKs) with properties suggestive of transmembrane sensors between the cell wall and the cytoplasm. Recent results show that WAKs are bound to pectin and are necessary for normal leaf cell enlargement and other growth processes.     

Plant biotechnology for lignocellulosic biofuel production

Lignocelluloses from plant cell walls are attractive resources for sustainable biofuel production. However, conversion of lignocellulose to biofuel is more expensive than other current technologies, due to the costs of chemical pretreatment and enzyme hydrolysis for cell wall deconstruction. Recalcitrance of cell walls to deconstruction has been reduced in many plant species by modifying plant cell walls through biotechnology. These results have been achieved by reducing lignin content and altering its composition and structure. Reduction of recalcitrance has also been achieved by manipulating hemicellulose biosynthesis and by overexpression of bacterial enzymes in plants to disrupt linkages in the lignin–carbohydrate complexes. These modified plants often have improved saccharification yield and higher ethanol production. Cell wall‐degrading (CWD) enzymes from bacteria and fungi have been expressed at high levels in plants to increase the efficiency of saccharification compared with exogenous addition of cellulolytic enzymes. In planta expression of heat‐stable CWD enzymes from bacterial thermophiles has made autohydrolysis possible. Transgenic plants can be engineered to reduce recalcitrance without any yield penalty, indicating that successful cell wall modification can be achieved without impacting cell wall integrity or plant development. A more complete understanding of cell wall formation and structure should greatly improve lignocellulosic feedstocks and reduce the cost of biofuel production.     

Plant biotechnology Biochemical engineering

A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Biotechnology.     

Plant and algal cell walls: diversity and functionality

Background

Although plants and many algae (e.g. the Phaeophyceae, brown, and Rhodophyceae, red) are only very distantly related they are united in their possession of carbohydrate-rich cell walls, which are of integral importance being involved in many physiological processes. Furthermore, wall components have applications within food, fuel, pharmaceuticals, fibres (e.g. for textiles and paper) and building materials and have long been an active topic of research. As shown in the 27 papers in this Special Issue, as the major deposit of photosynthetically fixed carbon, and therefore energy investment, cell walls are of undisputed importance to the organisms that possess them, the photosynthetic eukaryotes (plants and algae). The complexities of cell wall components along with their interactions with the biotic and abiotic environment are becoming increasingly revealed.

Scope

The importance of plant and algal cell walls and their individual components to the function and survival of the organism, and for a number of industrial applications, are illustrated by the breadth of topics covered in this issue, which includes papers concentrating on various plants and algae, developmental stages, organs, cell wall components, and techniques. Although we acknowledge that there are many alternative ways in which the papers could be categorized (and many would fit within several topics), we have organized them as follows: (1) cell wall biosynthesis and remodelling, (2) cell wall diversity, and (3) application of new technologies to cell walls. Finally, we will consider future directions within plant cell wall research. Expansion of the industrial uses of cell walls and potentially novel uses of cell wall components are both avenues likely to direct future research activities. Fundamentally, it is the continued progression from characterization (structure, metabolism, properties and localization) of individual cell wall components through to defining their roles in almost every aspect of plant and algal physiology that will present many of the major challenges in future cell wall research.     

Plant biotechnology in agriculture

Knowledge on plant genomes has progressed during the past few years. Two plant genomes, those of Arabidopsis thaliana and rice, have been sequenced. Our present knowledge of synteny also indicates that, despite plasticity contributing to the diversity of the plant genomes, the organization of genes is conserved within large sections of chromosomes. In parallel, novel plant transformation systems have been proposed, notably with regard to plastid transformation and the removal of selectable marker genes in transgenic plants. Furthermore, a number of recent works considerably widen the potential of plant biotechnology.     

Human lymphoid cell lines as targets for DRw

Summary Cultured human lymphoid cell lines (LCL) are useful as a source of target cells in several immunologic assays. More recently such cells have been used for the serological characterizations of the HLA-DR antigens. Typing of the same LCL in various laboratories during the VII Histocompatibility Workshop has given comparable results with a discordancy rate of less than 10%. This discordancy is likely to reflect the different sources of complement that can greatly alter the results of cytotoxic assays. The presence of naturally occuring antibody in rabbit complement to human cells can be avoided by: (a) absorbing with human cells at 0°C; (b) dilution with human serum; (c) dilution with heat-inactivated rabbit serum; (d) repeated freeze-thawing of the complement; or (e) careful selection of complement by screening procedures. Comparison of the results of HLA-DR typing of LCL with peripheral B-cells of the same donor show good correlations. However, LCL will occasionally give extra reactions perhaps due to the expression of new antigens. LCL can be coated with F(ab′)₂ fragments from antihuman β₂-microglobulin antibodies that block reactions of HLA-A, −B and −C antibodies allowing for discrimination of anti-DRw activity. Recipient of an American Heart Established Investigatorship. This work was supported by the Naval Medical Research and Development Command, Work Unit No. ZF51.524.013.1025, and by Grant Nos. AI 13154 AND CA 16071 from the National Institutes of Health.     

Plant biotechnology for food security and bioeconomy

This year is a special year for plant biotechnology. It was 30 years ago, on January 18 1983, one of the most important dates in the history of plant biotechnology, that three independent groups described Agrobacterium tumefaciens—mediated genetic transformation at the Miami Winter Symposium, leading to the production of normal, fertile transgenic plants (Bevan et al. in Nature 304:184–187, 1983; Fraley et al. in Proc Natl Acad Sci USA 80:4803–4807, 1983; Herrera-Estrella et al. in EMBO J 2:987–995, 1983; Vasil in Plant Cell Rep 27:1432–1440, 2008). Since then, plant biotechnology has rapidly advanced into a useful and valuable tool and has made a significant impact on crop production, development of a biotech industry and the bio-based economy worldwide.     

Cyclodextrins as a useful tool for bioconversions in plant cell biotechnology

The application of cyclodextrins as precursor solubilizers in biotechnological processes, in which plant cells are involved, is new. In this paper the possibilities for cyclodextrin facilitated bioconversions by freely suspended and/or immobilized plant cells or plant enzymes are demonstrated. After complexation with -cyclodextrin, the phenolic steroid 17-estradiol could be ortho-hydroxylated into a catechol, mainly 4-hydroxyestradiol, by a phenoloxidase from in vitro grown cells of Mucuna pruriens. By complexation with -cyclodextrin the solubility of the steroid increased from almost insoluble to 660 M. In addition, by complexation with -cyclodextrin, a solution of 3 mM coniferyl alcohol could be fed to cell cultures of Podophyllum hexandrum in order to enhance the accumulation of podophyllotoxin. Finally, the glucosylation of podophyllotoxin by cell cultures derived from Linum flavum was investigated. Four cyclodextrins: -cyclodextrin, -cyclodextrin, hydroxypropyl--cyclodextrin and dimethyl--cyclodextrin were used to improve the solubility of podophyllotoxin. Dimethyl--cyclodextrin met our needs the best and the solubility of podophyllotoxin could be enhanced from 0.15 to 1.92 mM. Podophyllotoxin--d-glucoside was formed at a rate of 0.51 mmol l^-1 suspension per day by the L. flavum cells growing in the presence of 1.35 mM podophyllotoxin, complexed with dimethyl--cyclodextrin.Abbreviations DW dry weight - E2 17-estradiol - FW fresh weight - PCV packed cell volume