Development of novel hop-derived bitter acid oxides with body fat-reducing effect

Transcription

1 Development of novel hop-derived bitter acid oxides with body fat-reducing effect 15 May th European Brewery Convention Takahiro Yamazaki 1, Yumie Morimoto-Kobayashi 1, Yoshimasa Taniguchi 2 (P009), and Mikio Katayama 1 1 Research Laboratories for Health Science & Food Technologies 2 Central Laboratories for Key Technologies

2 Hops are attractive materials with multiple functions Hops Fresh bitterness and flavor Multiple health benefits Relax effect as herbal medicines Gastrointestinal health as crude medicines Lots of studies have been performed about hops. Kirin Company, Limited 1

3 Body fat-reducing effect of iso-α-acids α-acids isomerization iso-α-acids Change of body fat area iso-α-acids Control 16mg 32mg 48mg Change of Total Fat Area (cm 2 ) 12-week ingestion Mean±SD, N = subjects/group Dunnett s post hoc test, *P<0.05, **P<0.01 Clinical Nutrition 2009, 28, 278 Application is limited due to its strong bitterness Kirin Company, Limited 2

4 Bitter acid oxides are generated during maturation process bitter acid oxides α-, β-acids Fresh hop pellets stored with air one year oxidation bitter acid oxides Matured hop pellets α-, β-acids Beer brewed with matured hop pellets little iso-α-acids and milder bitterness! Journal of the American Society of Brewing Chemists 1987, 45, 61 Do bitter acid oxides have health benefit like iso-α-acids? Hop material with mild bitterness and health benefit can be developed! Kirin Company, Limited 3

5 Contents 1. Chemical properties of matured hop 2. Health benefits on animals Details are presented by Dr. Taniguchi Poster No. P Mass production of matured hop 4. Health benefits on humans 5. Flavor Kirin Company, Limited 4

6 Reduction of maturation time Fresh hop pellets months 1 month Similar Peak Patterns Bitter acid oxides 60 2 days! Hop breed: Hallertau Perle J. Agric. Food. Chem. 2013, 61, Matured hop pellets can be prepared by heating at 60 C! Kirin Company, Limited 5

7 400 Generation pathways of bitter acid oxides Fresh hop pellets reverse phase HPLC detect: UV 270 nm 1a-c α-acids β-acids mau 200 2a-c 1a-c 2a-c hulupones 3a-c mau Matured hop pellets 7a 6a 5a 5b 5c 6b 7b 4a 4b 3a 4c 3b 3c humulinones 4a-c Journal of Agricultural and Food Chemistry 2013, 61, hydroxyallohumulinones tricyclooxyisohumulones A/B 6,7a-b Journal of Natural Products 2014, 77, Bitter acid oxides were generated from α-, β-acids 5a-c Kirin Company, Limited 6

8 mau Structural characteristics of bitter acid oxides Fresh hop pellets reverse phase HPLC detect: UV 270 nm α-acids β-acids β-tricarbonyl moiety 80 0 Matured hop pellets bitter acid oxides mau 40 β-tricarbonyl moiety 0 Journal of Agricultural and Food Chemistry 2013, 61, We defined α-, β-acid oxides with β-tricarbonyl moieties as matured hop bitter acids, MHBA Bioscience Biotechnology and Biochemistry 2015, 79, Kirin Company, Limited 7

9 uv min uv min Removal of α-, β-acids Matured hop pellets MHBA α-, β- acids Remained α-acids may cause bitterness via iso-α-acids hydrophilic hydrophobic water extract = elimination of α-, β-acids clarification concentration sterilization Extract MHBA α-, β- acids Relative bitter level MHBA were selectively extracted from matured hop pellets! Kirin Company, Limited 8

10 Contents 1. Chemical properties of matured hop 2. Health benefits on animals 3. Mass production of matured hop 4. Health benefits on humans 5. Flavor Kirin Company, Limited 9

11 Investigation of body fat-reducing effects of MHBA Control sample High fat diet Test sample High fat diet + MHBA 0.2% MHBA in high fat diet Normal 12 mice/group Test protocol 9 weeks body weight measurement Suppressed adiposity? adipose tissue weight measurement Kirin Company, Limited 10

12 MHBA have body fat-reducing effects Change of adipose tissue weight Change of body weight Mean±SE, t-test, **P<0.01 MHBA suppressed the increase in adipose tissue and body weights! Normal high fat diet + MHBA PloS one 2015, 10.6 Mean±SE, ANOVA with repeated measures, **P<0.01 Suppressed adiposity!! Kirin Company, Limited 11

13 How do MHBA show body fat-reducing effects? MHBA Expected mechanism BAT activation? fat burning body fat reduction SCIENCE, 2010, 328, 1113 Journal of Clinical Investigation, 2013, 123, 8, 3404 Thermogenic adipose tissue WAT (White adipose tissue) = generally known as fat BAT(Brown adipose tissue) = fat burning by thermogenesis Researches about anti-obesity focusing on BAT are getting attention around the world MHBA may induce fat burning by activating BAT, leading to body fat reduction Black area = Active BAT Kirin Company, Limited 12

14 MHBA induce thermogenesis in BAT Change of BAT temperature BAT temperature increased in MHBA group = Thermogenesis!! Mean±SD, N = 4 rats/group ANOVA with repeated measures, **P<0.01 PloS one 2015, other evidences demonstrating BAT was activated by MHBA MHBA induced fat burning by activating BAT! Kirin Company, Limited 13

15 Contents 1. Chemical properties of matured hop 2. Health benefits on animals 3. Mass production of MHBA-rich material 4. Health benefits on humans 5. Flavor Kirin Company, Limited 14

16 Mass production of MHBA-rich material Manufacturing process Fresh hop pellets Matured hop pellets Matured hop extract Mass production of matured hop extract Patents WO :HOPS OXIDATION PRODUCT, AND PRODUCTION METHOD AND USE THEREFOR WO :CARBONATED BEVERAGE CONTAINING HOPS OXIDATION REACTION PRODUCT EXTRACT WO :BEVERAGE CONTAINING AQUEOUS MEDIUM EXTRACT OF HOPS USED IN OXIDATION Kirin Company, Limited 15

17 Contents 1. Chemical properties of matured hop 2. Health benefits on animals 3. Mass production of MHBA-rich material 4. Health benefits on humans 5. Flavor Kirin Company, Limited 16

18 Evaluation of body fat-reducing effects on humans Control sample Test sample Beverage Beverage ml test beverage Matured hop extract 35 mg MHBA-containing matured hop extract 350 ml test beverage Test protocol one drink /day Measurement of body fat area and body fat ratio wks? Follow-up observation 25 BMI<30, n = 200 adult males and females Drastic changes in their diets and exercises were prohibited Kirin Company, Limited 17

19 Matured hop extract have body fat-reducing effects on humans Control 0 week 12 weeks Change of body fat area Change of body fat ratio Matured hop extract 0 week 12 weeks Pink: visceral fat area Blue: subcutaneous fat area Black and Gray: air or other tissues #P<0.05 vs control, **P<0.01 vs 0 week #P<0.05 vs control, *P<0.05 vs 0 week *No adverse events were detected Nutrition journal 2016, 15, 25 Matured hop extract safely reduced body fat on humans without changing their lifestyles!! Kirin Company, Limited 18

20 Contents 1. Chemical properties of matured hop 2. Health benefits on animals 3. Mass production of MHBA-rich material 4. Health benefits on humans 5. Flavor Kirin Company, Limited 19

21 Flavor of beverage with matured hop extract Preliminary sensory evaluation of beverage with/without matured hop extract was performed by three panelists Samples Control sample Test sample Beer-taste beverage Japanese brand PERFECT FREE Beer-taste beverage + matured hop extract Matured hop extract was added (60 ppm MHBA) Results No strong, but thick and complex bitterness No negative flavors Kirin Company, Limited 20

22 Summary 1st Stage Chemical properties of MHBA Stage Up! 2nd Stage Mass production of matured hop extract Stage Up! 3rd Stage Health benefits on humans Health benefits of MHBA on animals Flavor Kirin Company, Limited 21

23 Solution of social problems with matured hop extract Beer and food are the best combination! Diet-induced obesity Diabetes Hyperlipidemia Lifestyle-related disease Hypertension We developed novel hop material with body fat-reducing effects Matured hop extract Kirin Company, Limited 22

24 Future prospects of matured hop extract Matured hop extract Body fat-reducing effect Novel flavor Functional foods Applicable to Beers and beverages These products will conform to functional food system in Japan! Matured hop extract can make a healthy life with beers and beverages!! Kirin Company, Limited 23

25 Acknowledgements We d like to express our profound gratitude to the following people. Kazutoshi Shindo (Japan Women s University) Katsuya Nagai (ANBAS Co., Ltd. and Osaka University) Yoshihiro Ogawa (Kyushu University) Frank-Jürgen Methner (Technical University of Berlin) TTC Co., Ltd. Members of Shiga Factory (Kirin Brewery Co., Ltd.) Members of Research Laboratories for Alcoholic Beverage Technologies (Kirin Co., Ltd.) Kirin Company, Limited 24

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27 Quantitative analyses of MHBA in commercial beers Bioscience Biotechnology and Biochemistry 2015, 79, Kirin Company, Limited 26

28 Flavor of beer with matured hop extract Sensory evaluation of beer with/without matured hop extract was performed by three experienced panelists Samples Control Matured hop extract Beer Japanese brand KIRIN Ichiban Shibori Beer + matured hop extract Matured hop extract was added (60 ppm MHBA) Results KIRIN Ichiban Shibori No strong bitterness was detected, but thick and complex bitterness appeared No negative flavor related to oxidization was detected (oxidized flavor, cheese-like flavor) Type: pilsner type Ingredient: malt, hop Kirin Company, Limited 27