Our analytical laboratory is equipped with state-of-the-art equipment for accurate and efficient measurement of metabolites produced by the various microorganisms that are being studied for biofuels application, for characterizing chemical and physical structure of plant biomass material, and for the purification and characterization of important cell-wall-degrading enzymes. Some of the analytical platforms available include Fourier Transform Mid-Infrared Spectroscopy (FT-MIR), Fourier Transform Near-Infrared Spectroscopy (FT-NIR) (2), High Throughput Robotic Platform, liquid Chromatography Mass Spectroscopy (LC-MS), Gas Chromatography (GC), High Pressure Liquid Chromatography (HPLC) system, and Spectroscopic Plate Readers. These platforms are being used to assess chemical and structural changes to lignocellulosic materials as they move through different conversion processes, produce very pure component of labeled cellulases for our imaging work, and to measure the products of enzymatic or microbial conversion.
This laboratory primarily houses Dr. Larry Walker's undergraduate and graduate research assistants, and research support specialists. The equipment in this laboratory has been selected to provide quantitative measures of enzymatic and microbial activities. In addition, the laboratory is equipped for quantitative molecular ecology studies for a number of mixed-culture microbial conversion processes of interest to Dr. Walker and his research team. The Biomass Conversion Laboratory houses equipment for DNA and RNA analysis, such as a DNA-free safety cabinet, two PCR machines, gel electrophoresis apparatuses, a 24-capillary electrophoresis unit, a microplate reader with fluorescence capabilities, an imager system (G:box) both designed for fluorescent, luminescence and UV/Vis analysis, and a gas chromatograph. This laboratory also contains 3 incubators (ambient to 80°C), and is equipped for inorganic and organic chemistry. The laboratory is also equipped with an HPLC system equipped with a dual pump system, fluorescent, refractive index and UV/Vis online detectors, temperature oven, and auto-sampler, and is routinely used for soluble sugars analysis from biomass samples.
This office was created to provide a home away from home for researchers to have ready access to BRL facilities and to encourage multi-disciplinary dialogue and interactions. The office has fifteen cubicles with internet connections, a conference table and a small library. Three of the cubicles have high-end work stations for manipulating images, processing FT-IR spectrums, and for computationally intense data processing and analyses. The remaining cubicles are equipped with lap-top computers for use by BRL researchers. There is also a kitchenette with a microwave for warming food and a refrigerator.
Given the BRL focus on deploying industrial biotechnology for the development of robust biofuels and bioproducts systems we have developed a microbial conversion facility that allows researchers to carry our high-throughput screening of novel microorganisms, perform basic microbial growth and development studies, and to assess microbial kinetics in controlled fermentors at the 2, 14 and 150L scale. This laboratory houses 16 New Brunswick Scientific BioFlo 310 fermentation units ranging from 2.5L up to 16L, the fermentation lab is designed to allow for scale up. The lab consists of 4 large dry air incubators, CO2 incubators, 4 high temp water bath incubators, a biosafety cabinet, and an anaerobic chamber with built in incubators. Having such capacity allows for various techniques and a variety of experiments over a range of organisms from aerobes to anaerobes. Large volume fermentation can also be used for production of enzymes and optimization of fermentation processes.
The microscopy laboratory was designed with the objective of enabling the study of cellulose deconstruction from the micro- down to the nanoscale with high spatial and temporal resolution through the use of fluorescence microscopy. This laboratory houses three microscopes: an inspection microscope equipped with a color camera and dark field and phase contrast microscopy capabilities; a laser scanning confocal fluorescence microscope equipped with five laser excitation lines, motorized stage, and focus correction capabilities; and a total internal reflection fluorescence microscope equipped with three excitation laser lines, a high sensitivity EMCCD camera, motorized stage, and focus correction capabilities. This equipment enables researchers to probe the distributions of fluorescently labeled molecules in three dimensional structures, with submicron resolution, as well as to track individual fluorescent molecules with accuracies down to the tens of nanometers. In addition, existing heated stage and objective heaters allow the study of samples under controlled environmental and temperature conditions. Sample research projects currently being conducted using equipment in this laboratory are:
For plant and algae research we have two walk-in constant temperature growth chambers. These chambers can be set for a desired light intensity and light duration at a specified temperature. This ensures that experiments can be conducted under stable and reproducible conditions. Specialized equipment can be brought into the chambers to help provide mechanical mixing and/or nutrient delivery for specific growth experiments. Laboratory bench space behind the chambers is used for medium preparation and necessary monitoring equipment.
Given the need to extract biofuels and bioproducts at high concentrations BRL researchers are exploring high-solids conversion process with solids content of 20% to 50% solids wt/wt. This exploration includes understanding and modeling of key mass and energy transport processes, development of pure and mixed culture fermentation kinetics that are developed around the challenging microbial environment that is created during solid-state conversion, and integrating our understanding of the transport processes and microbial conversion kinetics to generate models that accurately simulate and predict process dynamics and product yields. Current work under ways is focused on exploring mixed culture population dynamics of the aerobic degradation of switch grass. This research activity is focused on exploring challenges to minimizing post-harvest losses from biomass storage. This laboratory houses a set of 50L static bed reactors, and several ½ L rotating drum reactors that can be operated in constant temperature incubators.
Producing a concentrated solution of monosaccharaides from biomass for subsequent microbial conversion to fuels and chemicals is a key bottleneck in the production of materials or fuels from biomass. Two pretreatment platforms have been developed for the BRL: alkaline pretreatment and bi-phasic H2O andCO2. Alkaline pretreatment is being conducted at multiple scales, from 1 to 20 grams at a time. Adequate mixing is imperative for alkaline pretreatment, and temperature–controlled incubator shakers in the pretreatment lab allow multiple pretreatment reactions to be run simultaneously at temperatures that range from 25 to 55 degrees Celsius. After solids separation, soluble samples are analyzed with HPLC, and solid samples are analyzed with FTIR–ATR spectroscopy. In addition, there is major capacity to carry–out biphasic pretreatment using unstirred and stirred reactors that are capable of processing 20 to 200 grams of materials. This laboratory is also equipped with a hammer mill for processing metric ton quantities and a small Wiley mill for processing analytical grade materials.
Discovery of novel cell-wall degrading enzymes (CWDEs), explorations on the molecular mechanisms of CWDEs and the development of more effective enzyme cocktails for the saccharification of plant cell-wall carbohydrates is the mission of this laboratory. In place is a Bio-Rad Biologic LP protein purification system for extracting enzymes of interest from the culture filtrate obtained from a diverse set native or cloned fungal and bacterial systems. There is an Eppendorf epMotion 5075 robotic platform for preparatory work for multiple 96-well plates. In addition, there is a robotic platform for high-throughput screening of CWDEs and mixtures of these enzymes. This robotic platform includes a Caliper Life Science Twister II robotic handler, a LiConic plate incubator, BioTEk plate-reader and a barcode tracking system. Current activities underway in the laboratory focus on identifying novel CWDEs from plant pathogenic fungi,boosting the activities of lignin-peroxidases, and assessing the degree of synergistic effects of enzyme cocktails.
The Research Support Lab houses major chemicals and vital supporting research equipment. Housed within the lab are 2 large volume nickel clad autoclaves (24x36x72) with sliding rack systems, 2 benchtop autoclaves, 2 large -80°C freezers, 2 large volume centrifuges with a max capacity of 6L each, large volume dishwasher, analytical balances, and a gas manifold system for nitrogen, oxygen, and carbon dioxide. The lab also contains an active fume hood for use of specialized applications. This laboratory is ideal for preparation of buffers, media, sterilization, and centrifugation work. The variety of equipment allows multiple users to multi-task and stay within the same laboratory.
