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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Healthy Processed Foods Research » Research » Research Project #438294

Research Project: Prevention of Obesity Related Metabolic Diseases by Bioactive Components of Food Processing Waste Byproducts and Mitigation of Food Allergies

Location: Healthy Processed Foods Research

2021 Annual Report

Objective 1: Resolve how novel single or in combination bioactive phytochemicals may enable the prevention of obesity and/or obesity related metabolic dysfunction in animal models. • Sub-objective 1A: Investigate the interactions of polyphenols on bile acids, proteins, and enzymes in intestinal lumen and their effect on intestinal permeability and FXR activation. • Sub-objective 1B: Evaluate ingredients containing two or more bioactive compounds such as polyphenols encapsulated for synergism in obese animal models. • Sub-objective 1C: Evaluate polyphenols encapsulated and/or immobilized in plant or microbial cell ghosts or gel bioactive fiber matrices such as HPMC, alginate, or glucomannan. Objective 2: Following gut fermentation of phytochemicals, determine the metabolites and/or peptide products that prevent inflammation in cell culture or animal models. • Sub-objective 2A: Identify major metabolites and proteins in extracts from food processing byproducts fermented by gut bacteria. • Sub-objective 2B: Culture 3T3-L1 adipocytes and lipopolysaccharide-activated RAW 264.7 macrophages with metabolites and/or peptides identified in 2A to assess anti-inflammatory response and lipid accumulation. Objective 3: Integrate measurable allergenic properties with methods to mitigate food allergens in nuts and dairy. • Sub-objective 3A: Develop methods for investigating conformational allergenic determinants of food allergens and assess their prevalence. • Sub-objective 3B: Define, characterize, and develop methods to detect novel nut allergens.

Hypothesis 1A: Polyphenols complexed by proteins modulate bile acid interactions with gut microbe. Specific research procedures: A high fat diet induced obesity (DIO) mouse (C57BL/6J) will be used. Polyphenols, conjugated and free bile acids in the intestinal lumen, colon, and feces will be measured by HPLC and intestinal permeability by FITC dextran. The expression of FGF15 and FXR expression will be measured by RT-PCR. Inflammatory and diabetic biomarkers by BioPlex multiplex ELISA. Hypothesis 1B: Synergistic interactions exist between polyphenols and other phytochemicals. Specific research procedures: The DIO mice will be fed single compounds or combinations. Polyvinylalcohol removal of polyphenolics and loss of activity would suggest synergistic compound is a polyphenol. Efflux pump inhibitors will be fed with grape seed proanthocyanidins and bile acids. Gut microbiome will be analyzed. Hypothesis 1C: Polyphenols beneficial health effects may be due to reducing the rapid influx of sugar, fat and other nutrients from rapidly digested foods that overload organs by binding to and inhibiting digestive enzymes. Specific research procedures: Yeast, bacteria, and plant cell ghosts infused with grape seed polyphenols, resveratrol or other bioactive phytochemicals will be provided by UC Davis. The encapsulated polyphenols will be tested in the DIO model described in Objective 1A. Hypothesis 2A: Fermentation produces absorbable phytochemicals from plant materials and bacterial protein/peptides. Specific research procedures: Extracts of food processing byproducts from apples, red beets, eggplants, grapes, olives, and cereal brans will be fermented with B fragilis, L plantarum, C perfringens, Bifidobacterium longum. The extract will be analyzed for total protein content, nucleic acids, and total polyphenolics. Specific phytochemicals will be analyzed by LC/MS. Hypothesis 2B: Products of fermentation and/ or bacterial derived peptides inhibit inflammation or lipid accumulation in adipose tissue. Specific research procedures: Glucose metabolism and lipid accumulation of 3T3-L1adipocytes and inflammatory responses in RAW264.7 macrophages will be measured with or without additions of substances from 2A. Hypothesis 3A: Structural information can be used to develop a new method that is applicable to map conformational IGE epitopes. Specific research procedures: An alternative ELISA format would use His-tagged allergens immobilized to N2+ coated microtiter plates. Final reverse mutant will have a His-tag. The pTNS2 vector will be used to construct plasmids for making the C-terminal His-tagged ßLG and its mutant by omitting the stop codon. To map the conformational IgE epitopes in ßLG, sera from 20 subjects with IgE antibodies against milk allergens will be used. Hypothesis 3B: PRU DU 8 orthologs in other plants are also food allergens. Specific research procedures: To characterize Pru du 8, we will use the recombinant allergen as antigen and make polyclonal antibodies in rabbits commercially. We will isolate the coding sequence for the new protein from the corresponding tree nuts to deduce the translated protein sequence.

Progress Report
Research has continued on bioactive food components that prevent or reduce metabolic dysfunctions related to obesity in animal models. Most of the bioactive compounds of interest have low solubility or are insoluble in aqueous solutions such as the digestive system. This property reduces their bioavailability and direct uptake into the blood stream. Passing through the intestine, these bioactive compounds interact with the microbes in the lower gut, are metabolized into smaller compounds, and are easily absorbed into the host body. The bioactive compounds also alter the composition of the microbial families and it is not clear whether the metabolites or the changes in the distribution of the microbiota cause the observed physiological effects. In order to improve bioavailability, resveratrol, a model bioactive compound, was incorporated into grape skin cells or yeast cells, in order to reduce crystals or solid resveratrol from forming. This maximizes their bioavailability, yet protects them from the acid environment of the stomach. The cells carrying resveratrol were introduced into the stomach of mice, and fluorescence was used to follow resveratrol in the digestive system. It was found that most of the resveratrol was retained in the stomach cells but some was released in the small intestine. Most of the resveratrol was delivered into the cecum and colon. These studies suggest that natural plant materials can be used to deliver bioactive molecules to gut microbes and increase their biological effects due to increased bioavailability.

1. Black and white sorghum brans reduce weight gain in mice on high fat diets. ARS scientists, in Albany, California, showed that the bran component of sorghum grains have health benefits. Sorghum brans contain high levels of bioactive polyphenolics different from other grains. Their research showed that mice on high fat diets supplemented with sorghum brans had reduced risk for obesity and related metabolic diseases. The extracted polyphenolics from white bran and the fiber fraction from the black bran were most effective at decreasing weight gain. The United States is the world’s largest producer of sorghum and this research suggests increasing its consumption would benefit human health and farm economies.

Review Publications
Ahammed, S., Liu, F., Wu, J., Khin, M., Yokoyama, W.H., Zhong, F. 2021. Effect of transglutaminase crosslinking on solubility property and mechanical strength of gelatin-zein nanocomposite films. Food Hydrocolloids. 116. Article 106649.
Alves, P.L., Berrios, J.D., Pan, J., Yokoyama, W.H. 2020. Black, pinto and white beans lower hepatic lipids in hamsters fed high fat diets by excretion of bile acids. Food Production, Processing, and Nutrition. 2. Article 25.
Chen, L., Yokoyama, W.H., Liang, R., Zhong, F. 2019. Enzymatic degradation and bioaccessibility of protein encapsulated ß-carotene nano-emulsions during in vitro gastro-intestinal digestion. Food Hydrocolloids. 100. Article 105177.
Guo, L., Yokoyama, W.H., Chen, M., Zhong, F. 2021. Konjac glucomannan molecular and rheological properties that delay gastric emptying and improve the regulation of appetite. Food Hydrocolloids. 120. Article 106894.
Ahammed, S., Liu, F., Khin, M., Yokoyama, W.H., Zhong, F. 2020. Improvement of water resistance and ductility of gelatin film by zein. Food Hydrocolloids. 105. Article 105804.
Liu, B., Fang, Z., Yokoyama, W.H., Huang, D., Zhu, S., Li, Y. 2020. Interactions in starch co-gelatinized with phenolic compound systems: effect of complexity of phenolic compounds and amylose content of starch. Carbohydrate Polymers. 247. Article 116667.
Seo, K., Kim, D., Yokoyama, W.H., Kim, H. 2020. Synbiotic effect of whole grape seed flour and newly isolated kefir lactic acid bacteria on intestinal microbiota of diet-induced obese mice. Journal of Agricultural and Food Chemistry. 68(46):13131-13137.
Chen, L., Liang, R., Yokoyama, W.H., Alves, P., Pan, J., Zhong, F. 2020. Effect of the co-existing and excipient oil on the bioaccessibility of ß-carotene loaded oil-free nanoparticles. Food Hydrocolloids. 106. Article 105847.
Rai, R., Merrell, C., Yokoyama, W.H., Nitin, N. 2020. Infusion of trans-resveratrol in micron-scale grape skin powder for enhanced stability and bioaccessibility. Food Chemistry. 340. Article 127894.
Wang, B., Zhe, Y., Yokoyama, W.H., Chiou, B., Chen, M., Lui, F., Zhong, F. 2021. Collagen peptides with DPP-IV inhibitory activity from sheep skin and their stability to in vitro gastrointestinal digestion. Food and Function. 42. Article 101161.
Villanueva-Suarez, M., Mateos-Aparicio, I., Perez-Cozar, M., Yokoyama, W.H., Redondo-Cuenca, A. 2019. Hypolipidemic effects of dietary fibre from an artichoke by-product in Syrian hamsters. Journal of Functional Foods. 56:156-162.
Chen, X., Liang, R., Zhong, F., Yokoyama, W.H. 2020. Effect of beta-carotene status in microcapsules on its in vivo bioefficacy and in vitro bioaccessibility. Food Hydrocolloids. 106. Article 105848.
Lui, B., Zhu, S., Zhong, F., Yokoyama, W.H., Huang, D., Li, Y. 2021. Modulating storage stability of binary gel by adjusting the ratios of starch and kappa-carrageenan. Carbohydrate Polymers. 268. Article 118264.
Fan, X., Wang, Y., Guo, F., Zhang, Y., Jin, T. 2020. Atomic-resolution structures of type I ribosome inactivating protein alpha-momorharin with different substrate analogs. International Journal of Biological Macromolecules. 164:265-276.
Li, C., You, Y., Chen, D., Gu, Z., Zhang, Y., Holler, T.P., Ban, X., Hong, Y., Cheng, L., Li, Z. 2020. A systematic review of rice noodles: raw material, processing method and quality improvement. Food Chemistry. 107:389-400.
Li, C., Ban, X., Zhang, Y., Gu, Z., Hong, Y., Cheng, L., Li, Z. 2020. Rational design of disulfide bonds for enhancing the thermostability of the 1,4-a-glucan branching enzyme from geobacillus thermoglucosidasius stb02. Journal of Agricultural and Food Chemistry. 68(47):13791-13797.
Li, C., You, Y., Zhang, Y., Xie, X., Xu, Q., Gu, Z., Ban, X., Tang, X., Hong, Y., Cheng, L., Li, Z. 2021. Maltose binding site 2 mutations affect product inhibition of Bacilluscirculans stb01 cyclodextrin glycosyltransferase. International Journal of Biological Macromolecules. 175:254-261.
Friedman, M., Tam, C.C., Cheng, L.W., Land, K.M. 2020. Anti-trichomonad activities of different compounds from foods, marine products, and medicinal plants: a review. BMC Complementary Medicine and Therapies. 20:271.
Friedman, M., Tam, C.C., Kim, J., Escobar, S., Gong, S., Liu, M., Yu Mao, X., Do, C., Kuang, I., Boateng, K., Ha, J., Tran, M., Alluri, S., Le, T., Leong, R., Cheng, L.W., Land, K.M. 2021. Anti-parasitic activity of cherry tomato peel powders. Foods. 10(2). Article 230.
Friedman, M., Xu, A., Lee, R., Nguyen, D.N., Phan, T.A., Hamada, S.M., Panchel, R., Tam, C.C., Kim, J., Cheng, L.W., Land, K.M. 2020. The inhibitory activity of anthraquinones against pathogenic protozoa, bacteria, and fungi and the relationship to structure. Molecules. 25(13):3101.
Crawford, L.M., Kahlon, T.S., Chiu, M.M., Wang, S.C., Friedman, M. 2019. Acrylamide content of experimental and commercial flatbreads. Journal of Food Science. 84(3):659-666.
Kim, J., Cheng, L.W., Chan, K.L., Tam, C.C., Mahoney, N.E., Friedman, M., Shilman, M.M., Land, K.M. 2020. Antifungal drug repurposing. Antibiotics. 9(11):812.
Chen, L., Yokoyama, W.H., Alves, P., Tan, Y., Pan, J., Zhong, F. 2021. Effect of encapsulation on ß-carotene absorption and metabolism in mice. Food Hydrocolloids. 121. Article 107009.
Tan, Y., Tam, C.C., Rolston, M., Alves, P., Chen, L., Meng, S., Hong, H., Chang, S.K., Yokoyama, W.H. 2021. Quercetin ameliorates insulin resistance and restores gut microbiome in mice on high fat diets. Antioxidants. 10(8). Article 1251.
Tan, Y., Tam, C.C., Meng, S., Zhang, Y., Alves, P., Yokoyama, W.H. 2021. Cooked black turtle beans ameliorate insulin resistance and restore gut microbiota in C57BL/6J mice on high-fat diets. Foods. 10(8). Article 1691.
Chen, F., Li, H., Fan, X., Li, Y., Zhang, C., Zhu, L., Hu, J., Kombe Kombe, A., Xie, J., Yin, D., Zhang, Y., Sun, J., Tang, R., Jin, T. 2021. Identification of a novel major allergen in buckwheat seeds: fag t 6. Journal of Agricultural and Food Chemistry.
Arellano, S., Law, B., Friedman, M., Ravishankar, S. 2021. Essential oil microemulsions inactivate antibiotic-resistant Salmonella Newport and spoilage bacterium Lactobacillus casei on Iceberg lettuce during 28-day storage at 4°C. Food Control. 130. Article 108209.
Chen, C.H., Marchello, J., Friedman, M., Ravishankar, S. 2021. Plant extracts and essential oils at concentrations acceptable to a sensory panel inactivate salmonella typhimurium dt104 in ground pork. Food and Nutrition Sciences. 12(2):162-175.