1 Capsaicinoids: A ReviewShinsuke Agehara, Ryan Barnes Laura Masor Andrea Roque Fall 2013
2 Presentation Outline Capsaicinoids Chemistry- RyanApplication to Medicine/Health- Andrea Breeding & Genetics- Laura Environmental effects- Shinsuke
3 Introduction to CapsaicinoidsGroup of phytochemicals only produced in genus Capsicum peppers (jalapeno, habanero, bell) Makes peppers hot Widespread consumption of capsaicinoids Purposed multiple benefits to health
4 Capsaicinoid SynthesisSynthesized in placenta of pepper (Curry et al., 1999)
5 Capsaicin and Dihydryocapsaicin(Schweiggert et al., 2006).
6 Capsaicin and Dihydryocapsaicin(Schweiggert et al., 2006).
7 Scoville Heat Units Invented by William Scoville in 1912.A sample is diluted until pungency cannot be detected. No Capsaicinoids ( 0 SHU) 1 ppm Capsaicin ~ 15 SHU
8 Capsaicinoids SHU Capsaicin ~ 16,000,000 SHUDihydrocapsaicin ~ 15,000,000 SHU Jalapeno – 2,500 – 10,000 SHU
9 Pungency Capsaicinoids cause a painful sensation when contacted by mammals. Transient Receptor Potential Vanilloid subfamily member 1 (TRPV1) also known as the capsaicin or vanilloid receptor
10 TRPV1 TRPV1 provides the sensation of scalding heat and pain.TRPV1 receptors are activated by: Heat Protons Calcium Capsaicinoids
11 TRPV1
12 TRPV1 Repeated activation of TRPV1 lowers pain responsiveness.Balance of extra/intra cellular Ca2+ Allows body to adapt to sustained stimuli over time.
13 Capsaicinoid ToxicityCapsaicin LD50 in rats reported at ~ 150 mg/Kg Human LD50 is ~ g/kg Symptoms of acute exposure: Skin Irritation Blindness/Lacrimation (Tears) Burning Sensation
14 Capsaicinoids: Medicine and HealthAnalgesic Effects Anticarcinogenic Effects Treatment for obesity and obesity-related illnesses
15 Pharmacokinetics Cmax = 2.47 + 0.13 ng/ml Tmax = 47.08 + 1.99 mint ½ = min
16 Analgesic Effect Capsaicin patches and topical creamstopical creams % capsaicin, FDA. Qutenza - 8% capsaicin patch Adlea - capsaicin injection long-term relief for osteoarthritis, post- surgery and neuropathic pain
17 Mode of Action Capsaicin induces initial period of heightened sensitivity, followed by a decrease in sensitivity Releases substance P from sensory nervous fibers Attaches to TRPV1, inducing an inflammatory response. Frequent application of capsaicin leads to the depletion of substance P → analgesic effect
18 Anti-cancer PropertiesInduces apoptosis Blocks NF-kB, AP-1, and STAT3 Scavenge carcinogenic substances such as nitrosamine.
19 Contradictory EvidenceMouse study - Small cell lung cancer Decreased proliferation and viability. Prevents DNA damage. Rat study - drinking water with .002% capsaicin Developed diethylnitrosamine-initiated, enzyme-altered foci Rat study - diet with 10% chilies gastric and liver cancer
20 Obesity hyperglycemia, hyperinsulinemia, and hyperlipidemiaanti-inflammatory characteristics and β-adrenergic activity Rat studies decrease in fasting glucose, insulin, and triglyceride levels IL-6 conclusion: decrease due to upregulation of adiponectin, which regulates glucose levels and fatty acid breakdown
21 Breeding/Genetics of CapsaicinoidsBreeding peppers Important horticultural & agronomic traits, with capsaicinoids in mind For direct consumption fresh, dried, processed High Capsaicinoid for extraction medical/pharmaceutical/research
22 1st thing a breeder needs: A pool of germplasm Many genotypes available for utilization C. annuum- bell type- 0 SHU C. Chinense- habanero C. frutesecens- tabasco C. Chinense x C. frutesecens- ‘Bhut Jolokia’ >1 million SHU
23 The genetics of pungency Major gene controlC/Pun1 locus Dominant homozygous dom. CC or heterozygous Cc=hot homozygous recessive cc= mild Many modifier genes cap, cap7.2
24 Correlations between capsaicinoidsCapsaicin & dihydrocapsaicin Capsaicin & nordihydrocapsaicin
25 F1 exhibits heterosis/overdominance Generation means F1 exhibits heterosis/overdominance progeny mean higher than parental F2 and F3 populations transgressive segregation progeny segregates with some having lower mean than parental
26 Environmental EffectsShinsuke Agehara G x E interactions Environmental factors Summary
27 G x E Interactions 6 genotypes 3 locations in TX
28 Temperature Optimal in vitro enzyme activity = 37°CMajor factor on photosynthesis (glucose → valine → capsaicin) Type Treatment Capsaicinoid Yield Chili (mild) 24 → 30°C ↗ (53%) ↗ 28/16 → 28/28°C NS
29 Light Major factor on photosynthesis (glucose → valine → capsaicin)High absorption of light at 450 and 680 nm Type Treatment Capsaicinoid Yield Chili (mild) 18 → 24 h day ↗ NS Chili (hot) Blue and red LED ↗ (67%)
30 Nitrogen Capsaicin requires 3 nitrogenous moleculesMore N = chlorophyll ↗ = photosynthesis ↗ Type Treatment Capsaicinoid Yield Jalapeno 1 → 22.5 mM ↗ (x2) ↗ Habanero 1 → 15 mM ↗ (x3) NS
31 Other environmental factorsCapsaicinoid Yield Water stress ↗ or NS ↘ Elevation ↗ ↗ Potassium ↗ NS
32 Summary of Enviromental EffectsTo increase capsaicinoid production or control pungency at the desired level… Selection of appropriate cultivars for specific environmental conditions Manipulation of temperature, LED, and N fertilization
33 References Butcher, J.D., K.M. Crosby, K.S. Yoo, B.S. Patil, A.M.H. Ibrahim, D.I. Leskovar, and J.L. Jifon, Environmental and genotypic variation of capsaicinoid and flavonoid concentrations in Habanero (capsicum Chinense) peppers. HortScience 47: Blum, E., M. Mazourek, M. O'Connell, J. Curry, T. Thorup, K. Liu, M. Jahn, and I. Paran, Molecular mapping of capsaicinoid biosynthesis genes and quantitative trait loci analysis for capsaicinoid Bosland, P.W. and J.B. Baral, ‘Bhut Jolokia’—The World's Hottest Known Chile Pepper is a Putative Naturally Occurring Interspecific Hybrid. HortScience 42: Caterina, M. J.; Schumacher, M. A.; Tominaga, M.; Rosen, T. A.; Levine, J. D.; Julius, D., The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 1997, 389, Curry, J., M. Aluru, M. Mendoza, J. Nevarez, M. Melendrez, and M.A. O’Connell, Transcripts for possible capsaicinoid biosynthetic genes are differentially accumulated in pungent and non-pungent Capsicum spp. Plant Science 148:47-57. Gangadhar, B.H., R.K. Mishra, G. Pandian, and S.W. Park, Comparative study of color, pungency, and biochemical composition in chili pepper (Capsicum annuum) under different light-emitting diode treatments. HortScience 47: Gurung, T., S. Techawongstien, B. Suriharn, and S. Techawongstien, Impact of environments on the accumulation of capsaicinoids in Capsicum spp. HortScience 46: Johnson, T.S., G.A. Ravishankar, and L.V. Venkataraman, In vitro capsaicin production by immobilized cells and placental tissues of Capsicum annuum L. grown in liquid medium. Plant Science 70: Lewis Sr., R. J. Sax’s Dangerous Properties of Industrial Materials, 10th ed.; John Wiley & Sons, Inc.: New York, 2000; p 702 Monforte-González, M., A. Guzmán-Antonio, F. Uuh-Chim, and F. Vázquez-Flota, Capsaicin accumulation is related to nitrate content in placentas of habanero peppers (Capsicum chinense Jacq.). Journal of the Science of Food and Agriculture 90:
34 References Murakami, K., M. Ido, and M. Masuda, Fruit pungency of 'Shishito' pepper as affected by a dark interval in continuous fluorescent illumination with temperature alteration. J. Soc. High Tech. Agric. 18: Narasimha Prasad, B.C., V. Kumar, H.B. Gururaj, R. Parimalan, P. Giridhar, and G.A. Ravishankar, Characterization of capsaicin synthase and identification of its gene (cys1) for pungency factor capsaicin in pepper (Capsicum sp.). Proceedings of the National Academy of Sciences of the United States of America 103: Pingle, S. C.; Matta, J. A.; Ahern, G. P., Capsaicin Receptor: TRPV1 A Promiscuous TRP Channel. In Transient Receptor Potential (TRP) Channels, Flockerzi, V.; Nilius, B., Eds. Springer Berlin Heidelberg: 2007; Vol. 179, pp Radovich, T., K. Crosby, G. Teves, A. Arakaki, and A. Ahmad, It's Hot in Hawai'i: Capsacin Content of Hawaii-grown Chili Peppers, Hanai'Ai/The Food Provider. Rahman, M.J. and H. Inden, Effect of nutrient solution and temperature on capsaicin content and yield contributing characteristics in six sweet pepper (Capsicum annuum L.) cultivars. Journal of Food, Agriculture and Environment 10: Rosenbaum, T.; Gordon-Shaag, A.; Munari, M.; Gordon, S. E., Ca2+/Calmodulin Modulates TRPV1 Activation by Capsaicin. The Journal of General Physiology 2004, 123, Saito, A.; Yamamoto, M. Acute Oral Toxicity of Capsaicin in Mice and Rats. J. Toxicol. Sci. 1996, 21, Schweiggert, U., R. Carle, and A. Schieber, Characterization of major and minor capsaicinoids and related compounds in chili pods (Capsicum frutescens L.) by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. Anal. Chim. Acta 557:
35 References Sung, Y., Y.Y. Chang, and N.L. Ting, Capsaicin biosynthesis in water-stressed hot pepper fruits. Botanical Bulletin of Academia Sinica 46:35-42. Stewart, C., B.-C. Kang, K. Liu, M. Mazourek, S.L. Moore, E.Y. Yoo, B.-D. Kim, I. Paran, and M.M. Jahn, The Pun1 gene for pungency in pepper encodes a putative acyltransferase. The Plant Journal 42: Zewdie, Y. and P.W. Bosland, Capsaicinoid inheritance in an interspecific hybridization of Capsicum annuum × C. chinense. J. Amer. Soc. Hort. Sci. 125: by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. Anal. Chim. Acta 557:
36 Capsaicinoids: A ReviewQuestions? Capsaicinoids: A Review