THE SCIENCE AND ENGINEERING FOR A BIOBASED INDUSTRY AND ECONOMY

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1017973
• Multistate No.: S-OLD 1075
• Project Start Date: Oct 4, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Recipient Organization
CLEMSON UNIVERSITY
(N/A)
CLEMSON,SC 29634

Performing Department
Biosystems Engineering

Non Technical Summary
Cottonseed oil, a relatively minor vegetable oil, is of great interest due to its natural antioxidant composition, mainly from endogenous gossypol observed to enhance the life of biodiesel from which it was derived (Moser, 2012). A major concern of producers is enhancing the life of biodiesel to increase its selling capacity. Many researchers add antioxidants in the post production phase, but with gossypol being naturally present it may become a strong contender in the market. Since the crop cannot be used for food because gossypol is toxic, cottonseed oil containing gossypol does not directly compete with food crops (De Sousa, De Moura, De Oliveira, & De Moura, 2014; Dunn, 2005; Wang, Howell, Chen, Yin, & Jiang, 2009). Glandless cottonseed oil not containing gossypol has a visual appearance like that of other vegetable oils and may be used in food. This oil was compared to measure the effect of gossypol for conversion in terms of possible inhibition.Novozymes Inc. recently released Eversa Transform® as a liquid lipase. Several optimal parameters were recommended in the user handbook. The purpose of this investigation was to determine how influentialmethanol:oil molar ratio, water content and lipase content are to the conversion of biodiesel from cottonseed oil. This examination will also provide data that determines how effective Eversa Transform® is at converting fringe feedstocks as opposed to the recommended waste cooking and refined soybean oils.

Animal Health Component

33%

Research Effort Categories

Basic

33%

Applied

33%

Developmental

34%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
402	1719	2020	100%

Knowledge Area
402 - Engineering Systems and Equipment;

Subject Of Investigation
1719 - Cotton, other;

Field Of Science
2020 - Engineering;

Keywords

biodiesel

enzymes

biosystems

Goals / Objectives
Research and develop technically feasible, economically viable and environmentally sustainable technologies to convert biomass resources into chemicals, energy, materials in a biorefinery methodology including developing co-products to enable greater commercialization potential. Perform system analysis to support and inform development of sustainable multiple product streams (chemicals, energy, and materials) and use the insights from the systems analysis to guide research and policy decisions

Project Methods
The glandless cottonseed oil was treated with a full 2x3x3 factorial design varying water dosage, lipase dosage and temperature, respectively. The crude or heavy pigment cottonseed oil was treated with a 2x3 full factorial design varying only water and lipase content. Both oils of 100 mL each were placed into 250-mL Erlenmeyer flasks with a prescribed amount of lipase, water and methanol:oil ratio. The reaction occurred in a horizontal shaker at 250 rpm for 24 hours at 25, 35, or 45 °C. After the reaction was complete, the flasks were removed from the shaker and the fuel was washed with distilled water, before waiting another 24 hours for the fuel to separate. The washing removed the enzyme, excess glycerol and thus halted the reaction. Once washed, the samples were analyzed for FAME content.The amount of free and total glycerol was calculated using the ASTM D6584 method (ASTM International, West Conshohocken, PA, USA). The sample was analyzed by gas chromatography after silyating with N-methyl-N-trimethylsilyltrifluoracetamide (MSTFA). Calibration was achieved using two internal standards and four reference materials. The column operation was conducted using helium as a carrier gas with a flow rate of 3 mL/min. Column temperature was as follows: 50 °C for 1 min, followed by increases of 15 °C /min to 180 °C, 7 °C /min to 230 °C, and 10 °C /min to 380 °C, kept for 8 min.Determining the difference between the two oils conducted comparing 3x3 full factorial blocks, modifying water and lipase dosage. The two different species of cottonseed oil were compared using the mean mass conversion of the oil to determine the difference in lipase performance based on the oil differences. Induction period was measured using a model 732 Rancimat instrument (Metrohm, Herisau, Switzerland) following EN 15751. The induction period was the mean of two or three measurements depending on the samples tested.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Lignocellulosic biomass is a promising alternative feedstocksto the crude oilfor biofuel production through thermochemical and/or biological pathways. Agricultural residuals and energy crops often contain high contents of alkali metals, chlorine, silica, and other elements that promote slagging, fouling, corrosion, and gas emission during thermochemical conversion (e.g., combustion and gasification). Water leaching is a common method, but not always effective to reduce theseelements. Bioleaching by adding microbes to water leaching was introduced to leach cellulosic biomass in this study. Three microbial species including two fungi (Fusarium oxysporum and Aspergillus niger) and one bacterium (Burkholderia fungorum) were selected to leach four biomass feedstocks includingcorn stover, wheat straw, switchgrass, and sorghum. Among thethree microbes, A. niger was found to be themost efficient to removeelements by as much as980% in 48 h, and sorghum was relatively more amenable to bioleaching. With A. niger, the bioleaching with water to feedstock (w/w) ratio of 25 for 6 h was sufficient to leach K (85%), Cl (90%), Mg (60%), and P (70%) from sorghum. Overall, bioleaching is more efficient than water leaching except for Na. Studies on bioleaching mechanism indicated that the acidification resultingfrom organic acids produced by A. niger during bioleaching might contribute to the higher leaching efficiency over other microbial species and water leaching. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Biosystems Engineering researchers and students from multiple disciplines interested in biofuels. Biofuel producers including farmers of feedstock materials and biofuel production facilities. How have the results been disseminated to communities of interest?Publication in a journal. What do you plan to do during the next reporting period to accomplish the goals?Among three microbes, A. niger was found the most efficient to remove most elements by 80% in 48 h, and sorghum was relatively more amenable to bioleaching. With A. niger, the bioleaching with water to feedstock (w/w) ratio of 25 for 6 h was sufficient to leach K (85%), Cl (90%), Mg (60%), and P (70%) from sorghum. Overall, bioleaching is more efficient than water leaching except for Na. Studies on bioleaching mechanism indicated that the acidification resulted from organic acids produced by A. niger during bioleaching might contribute to the higher leaching efficiency over other microbial species and water leaching. We plan to conduct more experimental runs using a multiple bioreactor system and disseminate results through a journal article submitted in 2021.

Impacts
What was accomplished under these goals? Lignocellulosic biomass is one of promising alternative feedstocks to the crude oil for biofuel production through thermochemical and/or biological pathways. Agricultural residuals and energy crops often contain high contents of alkali metals, chlorine, silica, and other elements that promote slagging, fouling, corrosion, and gas emission during thermochemical conversion (e.g., combustion and gasification). Water leaching is a common method, but not always effective to reduce all such elements. Bioleaching by adding microbes to water leaching was introduced to leach cellulosic biomass in this study. Three microbial species including two fungi (Fusarium oxysporum and Aspergillus niger) and one bacterium (Burkholderia fungorum) were selected to leach four biomass feedstocks such as corn stover, wheat straw, switchgrass, and sorghum.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Zhang N, L Wang, K Zhang, TH Walker, P Thy, B Jenkins, Y. Zheng, 2019. Pretreatment of Lignocellulosic Biomass Using Bioleaching to Reduce Inorganic Elements, Fuel, 246: 386-393.

Progress 10/04/18 to 09/30/19

Outputs
Target Audience:Biosystems Engineering students and students from multiple disciplines interested in biofuels. Also, biofuel producers including farmers of feedstock materials and biofuel production facilities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training opportunities include use of pilot biodiesel, gasification and compost for training students and personnel to experiment directly with university waste streams for bioenergy use. How have the results been disseminated to communities of interest?ASABE What do you plan to do during the next reporting period to accomplish the goals?Continue work on enzyme catalyzed biodiesel from hemp products and supercritical fluid extraction of hemp oils, andon bioleaching pretreatment of biomass.

Impacts
What was accomplished under these goals? Eversa Transform was used as an enzymatic catalyst to transform glandless and crude (heavy pigment) cottonseed oils into biodiesel. The oils were reacted with methanol at a 6:1 molar ratio with modified amounts of water, lipase, and temperature. Reactions were conducted in the presence of lipase and water at doses of 2, 5, and 8 wt% and 1, 3, and 6 wt%, respectively. Product composition and conversion were determined using the gas chromatography method of ASTM D6584. Oxidative stability was determined following EN 15751. The conversion to fatty acid methyl esters averaged 98.5% across all samples. Temperature had the most significant effect on conversion (p < 0.0035). Lipase and water dosages did not affect conversion, while each had an effect with temperature that was significant across the difference between 3 and 1 wt% water content and between 8 and 5 wt% enzyme content between the two temperatures (p = 0.0018 and 0.0153), respectively. Induction periods (oxidative stability) of the glandless and crude cottonseed oils were significantly different, but there was no difference between the two oil conversions based on oil type.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Anderson ST, BR Moser, CM Drapcho, Y Zheng, TH Walker, 2019, Evaluation of dominant parameters in the performance of a lipase in the transesterification of cottonseed oil, Trans. ASABE, 62(2): 467-474. doi: 10.13031/trans.13003