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Executive Committee
HATSUMATA Shigeru HU Hong-ying
JUNG Jinyoung MATSUlYoshihiko NlSHlJlMAWataru ONO Yoshiro SHI Jianghong YEDLA Sudhakar Scientific Committee
WET2018 Committees
Ehime University Hokkaido University Kitasato University Kumamoto University Kyoto tJniversity Toyo University
Kanto Gakuin University
Center for Environment. Science in Saitama The University of Tokyo
National Institute of Technology, Nagaoka College Tohoku University
Ehime University
Niigata
Universityof
Pharmacyand
Applied SciencesMetawater Co Ltd Tshinghua University Yeungnam University Hokkaido University Hiroshima University
Kyoto Institute of Technology
South University of Science & Technology of China Indira Gandhi Institute of Development Research The University of Tokyo
Kyoto University Azabu University University Malaya
National lnstitute for Environmental Studies University of Wollongong
The University of Tokyo
Nagaoka University of Technology Osaka University
Gunma University Hanyang University
National Institute of Health Sciences United Nations University
National Institute of Technology, Gunma College Kyoto University
Chalmers University of Technology
National Institute of Technology, Ube College University of Yamanashi
Yokohama National University Kindai University
Tohoku University lwate University Organizing commiftee
Chair Vice Chair Commiftee member
Secretary Vice Secretary
Chief Secretary Committee member
HARUTA Shinsuke MATSUlYoshihiko SEI Kazunari HAMA Takehide HIDAKA Taira ISAKA Kazuichi KAMATA Motoyuki MISHIMA lori OGUMA Kumiko OSHIKI Mamoru SANO Daisuke WATANABE Kozo lGUCHlAkinori
SATOH Hiroyasu HlDAKATaira OHKOUCHlYumiko CHUAAdeline Seak Mav HAFUKAAkiTa
HAI Faisal lbney HASHIMOTO Takash HATAMOTO Masashi INOUE Daisuke ITO Tsukasa KIM Jong-Oh
KOBAYASHI Norihiro MASAGO Yoshifumi MIYAZATO Naoki MIZUNO Tadao MODIN Oskar NAKANO Yoichi NISHIDA Kei N|TTAMlTadashi OGATA Fumihiko TADA Chika
TERASAKI Masanori
Copyright: Those who wish to take a copy of this book are required to get permission from Japan Academic Association for Copyright Clearance (9-6-41 Akasaka Minato 107-0052 Tokyo, Japan Tel:
+81 -3-347 5-56 1 8 Emai | : i [email protected])
lssued on 14th July 2018
Japan Society on Water Environment Taiyo Bijutsu (Printing office)
WET2018 Program and Abstract
Green Plaza Fukagawatokiwa
201
2-9-7 Tokiwa Koto 135-0006 Tokyo, Japan +8'1-3-3632-53512-7-11
KiyosumiKoto
135-0024 Tokyo, Japan +81 -3-3642-6045Water and Environment Technology Conference 2018 (WET 2018)
Hydrogen Producing Ability of Extremely Halotolerant Bacteria from Salt-Damaged Soil in Thailand
Dyah Asri Handayani TAROEPRATJEKA* , **, Tsuyoshi IMAI*, Prapaipid CHAIRATTANAMANOKORN***, Alissara REUNGSANG****,*****
*Graduate School of Sciences and Technology for Innovation, Yamaguchi University, Yamaguchi 755-8611, Japan
**Department of Environmental Engineering, Institut Teknologi Nasional, Bandung, Jawa Barat 40124, Indonesia
***Department of Environmental Technology and Management, Faculty of Environment, Kasetsart University, Bangkok 10900, Thailand
****Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen 40002, Thailand
*****Research Group for Development of Microbial Hydrogen Production Process from Biomass, Khon Kaen University, Khon Kaen 40002, Thailand
3. Results
4A-11
1. Introduction
Growing Energy Demand
Fossil energy depletion
Climate change
& global warming
Urban air pollution
Development of alternative energy
(wind, solar, sun, biomass)
Waste sources
Bio- hydrogen
• Hydrogen gas has the highest energy density of common fuels expressed on a mass basis.
‘Next Generation
Industrial Biotechnology’
Cost reduction
Extremely halotolerant microorganism
• Fresh water
• Oxygen
• Sterilization
• Dark fermentation process for H 2 production
• Waste sources to provide sugars
Cultivation of fermentation
bottles
Experiment of hydrogen production:
Basic condition, different pH conditions, etc .
Process
development of H 2 production directly from cellulose waste
After substrate change, bottle is flushed with N
2gas to ensure anaerobic condition
500 mL serum bottle Liquid + solid volume ± 350 mL
Substrate is changed every ± 7 days for each bottle. Mixed liquor is taken out by glass syringe and centrifuged. The supernatant is removed, and pellets are mixed with new substrate
26% NaCl
Gas volume checked
everyday with glass syringe
• 75,100, 125 mL serum bottles
• substrate
• supernatant of fermentation bottles
Bottle filled with N
2gas to ensure anaerobic condition
Experiment serum bottles are kept in shaking water bath incubator in 35-37° C, 100 rpm Gas production checked every 6 to 24 hours with wetted glass syringe method
A (NH 4 ) 2 HPO 4 350 gram/L
NaHCO 3 (buffer) 2 gram/L K 2 HPO 4 (buffer) 2 gram/L
Yeast extract 0.0532 - 1 gram/L
Glucose 5 gram/L (fermentation bottles)
NaCl 3-10%, 15-26% w/v
(26% =351.35 gram/L) Trace elements A 2 mL/L
B 10 mL/L C 1 mL/L C CaCl 2 ·2H 2 O 150 gram/L B
KCl 75 gram/L NH 4 Cl 85 gram/L FeCl 3 ·6H 2 O 42 gram/L MgCl 2 ·6H 2 O 81 gram/L MgSO 4 ·7H 2 O 25 gram/L CoCl 2 ·6H 2 O 1.8 gram/L
• Gas volume is measured by wetted glass syringe method.
• Composition of biogas (H 2 ,N 2 , CH 4 and CO 2 ), was measured by gas chromatography (GC-8APT/TCD;
Shimadzu Corp. Japan) with activated charcoal 60/80 mesh column and Argon as carrier gas.
• Polymerase chain reaction-denaturing gel
electrophoresis (PCR-DGGE) was used to study
bacterial community structure differences before and after acclimatization.
Salt-damaged soil (Khon Kaen, Thailand)
Equations of fermentative reactions ∆G 0
C 6 H 12 O 6 + 4H 2 O → 2CH 3 COO − + 2HCO 3 − + 4H + + 4H 2 (1) -206 kJ C 6 H 12 O 6 + 2H 2 → CH 3 CH 2 2 COOH + 2CH 3 COO − + H + + 2H 2 (2) -254 kJ Standard Gibbs energy of formation for glucose fermentation
Equation (1): 1 mol of glucose will produce 4 mol of hydrogen 1 g of glucose STP conditions produce 498 mL of H 2 via the acetic acid (HAc) pathway Equation (2): 1 mol of glucose will produce 2 mol of hydrogen 1 g of glucose at STP conditions produce 249 mL of H 2 via the butyric acid (HBu) pathway
𝑯𝒚𝒅𝒓𝒐𝒈𝒆𝒏 𝒚𝒊𝒆𝒍𝒅 % = 𝑶𝒃𝒔𝒆𝒓𝒗𝒆𝒅 𝒄𝒖𝒎𝒖𝒍𝒂𝒕𝒊𝒗𝒆 𝒉𝒚𝒅𝒓𝒐𝒈𝒆𝒏 (𝒎𝑳)
𝑻𝒉𝒆𝒐𝒓𝒆𝒕𝒊𝒄𝒂𝒍 𝒄𝒖𝒎𝒖𝒍𝒂𝒕𝒊𝒗𝒆 𝒉𝒚𝒅𝒓𝒐𝒈𝒆𝒏 (𝒎𝑳) × 𝟏𝟎𝟎
Objectives:
• to investigate hydrogen production by extremely halotolerant bacteria
• to analyze bacterial community structure difference before and after acclimatization in saturated NaCl condition
2. Materials and Methods
0 10 20 30 40 50 60
0 1 2 3 4 5 6 7 8 9 10 11 12
Cumula tiv e H2 [mL]
Time [Day]
NaCl 15%
NaCl 20%
NaCl 26%
4. Conclusions
• After acclimatization, it was possible to produce biohydrogen under high salt concentration (26% w/v NaCl) with hydrogen yields of 0.66–1.15 mol H 2 /mol glucose .
• Biohydrogen production indicates that extremely halotolerant hydrogen-producing bacteria can exist under high salt concentrations.
• PCR-DGGE confirmed that changes in microbial community structure have occurred after 2 years of acclimatization in saturated NaCl condition.
Biohydrogen production at 3–10% salinity of salt-damaged soil from Khon Kaen Salt
concentration (%)
Biohydrogen production (ml)
Theoretical H 2 production (%) Yield
(molH 2 / mol glucose ) HAc pathway HBu pathway
3 10.9 14.7 29.5 0.61
3.5 10.9 14.7 29.5 0.61
5 9.46 12.8 34.6 0.53
7 13.4 18.1 36.2 0.75
7.5 7.43 10 20.1 0.41
10 18.1 24.5 49 1.01
Glucose 0.15 g (5,000 mg/L), inoculum 3,000 mg/L VSS, F/M ratio 1.5
Biohydrogen production at 15–26% salinity of salt-damaged soil from Khon Kaen Salt
Concentration (%)
Biohydrogen production (ml)
Theoretical H 2 production (%) Yield
(molH 2 / mol glucose ) HAc pathway HBu pathway
15 49.8 67.3 134.6 2.78
20 0.02 0.03 0.05 0.00
26 0 0 0 0.00
Glucose 0.15 g (5,000 mg/L), inoculum 3,000 mg/L VSS, F/M ratio 1.5
Cumulative hydrogen production before acclimatization
Soil Sample Biohydrogen production (ml)
Theoretical H 2 production (%) Yield
(molH 2 /mol glucose ) HAc pathway HBu pathway
Khon Kaen salt
damaged soil (1) 14.31 28.76 57.55 1.15
Khon Kaen salt
damaged soil (2) 8.22 16.53 33.07 0.66
Glucose 0.12 g (5,000 mg/L), inoculum 10 mL (3 mg/L VSS)
Biohydrogen production at 26% salinity after two years of acclimatization
0 2 4 6 8 10 12 14 16
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Cumula tiv e H2 [mL]
Time [Day]
Khon Kaen 1 Khon Kaen 2
Cumulative hydrogen production after 2 years of acclimatization
DGGE profiles of V3 region of 16s rDNA of acclimatized culture (A,C) and soil samples
of Khon Kaen (D,E,F)
A B C D E F G H I
** * * * * * * * ** * * * * * t * * * * * * * * * * * * * * tr * * * ** * 1. * * * Japan Society on Water Environment (JSWE)
Green-Plaza-Fukagawa-Tokiwa #20 I 2-9-7 Tokiwa, Koto-ku, Tokyo 135-0006 Japan
Tel. +8 l-3-36?2-535
I
Fax. +81-3-3632-5352 hnp://wwwj swe.orjp* * * * t *! * * * * * * * * * * * * * * * * * * * * + * * * * * 't! * * * * * * * * *
July l5th, 2018
This is to certifi that
Ms. Dyah Asri Taroepratjeka Yamaguchi University
had participated in the Water and Environment Tbchnolog,t Conference (WET20lB) fficially organized by Japan Society on Water Environment _fro* I4th to l5th July, 2018, held at Ehime (Jniversity (Bunlryo-cho 3, Matsuyama, Ehime, Japan),
and had presented the presentation entitled " Hydrogen
Producing Ability of Extremely Halotolerant Bacteria fro*
Salt-Damaged Soil in Thailand ".
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