講演要旨集

(1)

日時：2021年６月12日（土）13時00分開会

オンライン開催（広島大学大学院統合生命科学研究科）

日本農芸化学会中四国支部支部創立20周年記念

日本農芸化学会中四国支部第59回講演会

(2)

(3)

20

59

2021 6 12

10:00 11:00 Zoom

11:10 12:00 Zoom

13:00 13:15 2021 Zoom

13:15 13:55 Zoom

2021

Aryl Hydrocarbon Receptor Repressor

13:55 14:20 Zoom

2021

Thanutchaporn Kumrungsee

15:00 17:35 A D Zoom

−1−

(4)

Zoom

A1 A10 B1 B13 C1 C9 D1 D9

A1 A4 A8

A3 A7 A10 B1

B5 B9

B4 B8 B13 C1

C6

C5 C9 D1

D4 D7

D3 D6 D9 1.Zoom

2. 15:00

3. 9 2 1

−2 −

(5)

講演会

プログラム

(6)

(7)

20

59

2021 6 12

10:00 11:00 Zoom

11:10 12:00 Zoom

13:00 13:15 2021 Zoom

13:15 13:55 Zoom

2021

Aryl Hydrocarbon Receptor Repressor

13:55 14:20 Zoom

2021

Thanutchaporn Kumrungsee

15:00 17:35 A D Zoom

− 3 −

(8)

A-1 15:00 Aurantiochytrium

A-2 15:12 (bar)

1 1, 2

1 2

A-3 15:24 agm1 agm2

A-4 15:36 Cloning, overexpression and purification of a NADP-dependent malic enzyme with unusual thermostability from psychrophile Shewanella livingstonensis Ac10

1 1 1 1 1

1

A-5 15:48 D-

1 1

A-6 16:00 Thermus thermophilus c’

, ,

A-7 16:12 Thermotoga maritima

A-8 16-24 Thermus thermophilus HB8

− 4 −

(9)

A-9 16:36 Thermus thermophilus HB8

1, 2 1, 2 2 2

1 2

A-10 16:48 Purification and characterization of the membrane-bound aldehyde dehydrogenase AldFGH of Gluconacetobacter diazotrophicus PAL5.

Miah, R.¹, Nina S.¹, Kataoka, N.^1,2, Matsutani, M.³, Matsushita, K.^1,2, Yakushi, T.^1,2

1Grad. Sch. Sci. Tech. Innov.,²RCTMR, Yamaguchi Univ., ³Tokyo Univ. Agric

− 5 −

(10)

B-1 15:00 -

B-2 15:12 CoQ

1 2 2 2 2 1, 2, 3

1, 2, 3

1 2 3

B-3 15:24

1 2

B-4 15:36 COP

1 1

B-5 15:48

B-6 16:00 sde

B-7 16:12 Acidovorax cirulli Aave_4606

B-8 16:24 Aurantiochytrium

1 1

B-9 16:36

− 6 −

(11)

B-10 16:48 1- -3-

Pattana Kakumyan¹ ²

3 3

1 2 3

B-11 17:00 Eikenella corrodens

1 1

B-12 17:12 Ralstonia. solanacearum

Asmaa Ali Ahmed Ibrahim

B-13 17:24

− 7−

(12)

C-1 15:00

C-2 15:12 Nudix hydrolase

1 2

C-3 15:24

1 2

C-4 15:36

1 2

C-5 15:48

1 1 2

1 2

C-6 16:00 CPC

C-7 16:12 MYB

C-8 16-24 EGFR-ERK

1 1 2 1

1 2

− 8 −

(13)

C-9 16:36

1 1

1

− 9 −

(14)

D-1 15:00 STZ

Tolulope Peter Saliu¹ ¹ ¹

Thanutchaporn Kumrungsee¹ ¹

1

D-2 15:12 OGTT DPPIV AW WV

GLP-1

1 2 3

D-3 15:24

1 1 2 2

1 2

D-4 15:36 B12

1 1

D-5 15:48

D-6 16:00

Thanutchaporn Kumrungsee ¹

1

D-7 16:12

D-8 16-24

−10 −

(15)

D-9 16:36

−11−

(16)

(17)

受賞講演

講演要旨

(18)

(19)

2021

Aryl Hydrocarbon Receptor Repressor

Aryl Hydrocarbon Receptor AHR AHR Aryl Hydrocarbon Receptor Nuclear Translocator ARNT

Hypoxia Inducible Factor HIF HIF-2 HIF-1

2 ARNT HIF-1

AHR ARNT/HIF-1

AHR Aryl Hydrocarbon Receptor

Repressor AHRR HIF

AHRR HIF

Hypoxia Response Element AHRR

AHRR Single

Nucleotide Polymorphism SNP 1 Pro189Ala SNP

3013

52.5 47.5 51.47 BMI 23.38 kg/m

²

Estimated glomerular filtration rate eGFR 0.72 mg/dL 77.05 mL/min/1.73m

²

14.01 g/dL

58.7 26.5 14.7 AHRR

189 / Pro/Pro 1072 /

Pro/Ala 1296 / Ala/Ala 407

eGFR Ala

Ala

Ala AHRR

AHR HIF

9 8 AHRR

AHRR

0.08 mg/dL 0.52 mg/dL

AHRR

3 1

AHRR

AHRR AHR

−13 −

(20)

2021

EC 1.14.18.1

5-hydroxymethyl-2-furaldehyde cycloalliin B16 mRNA

cAMP

MITF B16

5-hydroxymethyl-2-furaldehyde cycloalliin cAMP mRNA

1,3-dihydroisobenzofuran-4,5,7-triol 6-hydroxy-L-tryptophan 1,3-dihydroisobenzofuran-4,5,7-triol

B16

mRNA

−14 −

(21)

2021

Thanutchaporn Kumrungsee

Ca

²⁺

Ca

²⁺

Trp-His His-Gly-Lys Trp-Val Trp-Ile

Ca

²⁺

Trp-His

Ca

²⁺

/ II CaMK II

Trp-His Ca

²⁺-CaM

CaM CaM

His

- -Ala

His

B

6

-

GABA

-His

B

6

B

6

-Ala

GABA

3R 3R

luciferase

imaging Serum amyloid

A3 luciferase

−15 −

(22)

(23)

一般講演

講演要旨

(24)

(25)

Aurantiochytrium

Aurantiochytrium 2

Aurantiochytrium limacinum SR21

NADPH NADPH

SR21 ALME 1

SR21 ALME

38 115 SR21

AMP-activated protein kinase ALSnf1-3 CRISPR-Cas9 ALSnf1-3

(bar)

1 1, 2 1 2

bar

S. pombe bar

bar

bar Cas9 RNA

S. cerevisiae bar

−17−

(26)

agm1 agm2

2

(Schizosaccharomyces pombe) 3

agm1 agm2 agm3 Agm3

Agm1 Agm2 Agm1 Agm2

agm pH 6.4 5 mM agm3

Agm3

Agm1 Agm2

agm3 agm1 agm2

Agm3 pH 6.4

5 mM agm2 agm3

Agm2

Cloning, overexpression and purification of a NADP-dependent malic enzyme with unusual thermostability from psychrophile Shewanella livingstonensis Ac10

1 1 1 1 1

1 1

NADP-dependent malic enzyme (NADP-ME) catalyzes a reversible oxidative decarboxylation of L-malate to pyruvate using NADP(H) as a cofactor with the help of Mg²⁺. It is widely distributed among many organisms, playing various roles. However, there are few reports on NADP-ME from pyschrophile even though it is an important provider of NADPH which is a vital ingredient for membrane phospholipids synthesis.

In the present research, a NADP-ME with unusual thermostability from psychrophile, Shewanella livingstonensis Ac10, was characterized. The active S. livingstonensis Ac10 NADP-ME formed 100 kD homodimer in solution. It was active at a very wide range of temperatures from 4°C to 80°C and remained stable at 60°C for 3 hours. Its thermostability was verified through circular dichroism measurement, the mid-point of thermal transition (Tm) value of S. livingstonensis Ac10 NADP-ME being 71.9°C, which was significantly higher than that of mesophilic counterpart, Escherichia coli MaeB (64.6°C). It was also found that S.

livingstonensis Ac10 NADP-ME had a more stable overall structure than the mesophilic one. Our results strongly suggested that not all proteins in psychrophile need to be cold-adapted. The fact that the psychrophilic NADP-ME was more active at higher temperatures also suggested that the NADP-ME may be involved in response to temperature changes.

−18 −

(27)

D

-

1 1

Streptomyces sp. 82F2 D- DAH Ser

DAH ^D- D-

DAH

Ac-D-Phe-OMe Trp

A267F DAH Cyclo(^D-Pro-^L-Arg)

D-Pro Arg-OEt ^D-Pro

A267F DAH

A267F DAH ^D-Pro-OBzl

D-Pro-OMe A267F DAH D-Pro-OBzl

Thermus thermophilus c

, ,

c

c TTCP c

TTCP M

s Bath c MCCP TTCP

X TTCP MCCP TTCP

MCCP Phe-13 Ile-38 TTCP

MCCP F13S I38V

CD

CD TTCP F13S I38V TTCP

TTCP TTCP

−19 −

(28)

Thermotoga maritima

AK -

Asa Asa

HseDH

AK HseDH AK/HseDH

AK

pET11a 80

20 DEAE 650 M Butyl 650 M

Superdex 200 pg 4

74.9 U/mg L-Lys L-Lys

Crystl screen 2-23 AK

Thermus thermophilus HB8

Thermus thermophilus HB8 Tth-PDI

S-S Tth-PDI PCR

(Protein Thermal Refolding)

sfGFP 2

sfGFP-2SS sfGFP-2SS DTT

sfGFP-2SS Tth-PDI pET39b

Tth-PDI His-tag

qRT-PCR sfGFP-2SS Tth-PDI

/ 70

60 25

sfGFP-2SS 12.5%

−20 −

(29)

Thermus thermophilus HB8

1, 2 1, 2 2 2

1 2

Arthrobacter globiformis M30

D- Thermus thermophilus HB8

T. thermophilus HB8

SDS-PAGE 73 kDa 150 kDa

T. thermophilus HB8

7.76×10^-2 U/mg 23.6

pH pH 9.5 pH 24 pH 7.0

85 10 90 1

70 L-

D- ^L- ^D- ^L- ^D- 5

10 Purification and characterization of the membrane-bound aldehyde dehydrogenase AldFGH of Gluconacetobacter diazotrophicus PAL5.

Miah, R.

¹

, Nina S.

¹

, Kataoka, N.

^1,2

, Matsutani, M.

³

, Matsushita, K.

^1,2

, Yakushi, T.

^1,2

(

¹

Grad. Sch. Sci. Tech. Innov.,

²

RCTMR, Yamaguchi Univ.,

³

Tokyo Univ. Agric)

Acetic acid fermentation is carried out by two-step oxidation of ethanol to acetic acid with the membrane-bound alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) of acetic acid bacteria. ALDH oxidizes acetaldehyde to acetic acid, coupled to the reduction of ubiquinone in the membrane. Then, the reducing equivalents link to the respiratory chain to reduce molecular oxygen to water. Structure and molecular mechanism of ALDH are not systematically studied yet. Therefore, some issues regarding the roles as well as chemical nature of prosthetic groups associated with ALDH remained a matter of debate (1). Recently, we reported that AldFGH, one of ALDHs, in Gluconacetobacter diazotrophicus PAL5 plays the major role in acetic acid fermentation and require a form of molybdopterin but not PQQ for ALDH activity through reverse genetic study (2).In the present study, we attempted to purify intact AldFGH complex using constructed triple deletion derivative aldSLC adhAB PQQ of PAL5 that had lost the genes for AldSLC an ALDH, ADH, and the biosynthetic pathway of pyrroloquinoline quinone for biochemical analysis. Previous report failed to detect ubiquinone reduction activity of purified ALDH from wild-type PAL5 strain with acetaldehyde (1) but we succeeded to detect it using ubiquinone-2 (Q2) that is a short variant of artificial ubiquinone analog. We will attempt to identify the prosthetic group of ALDH and study molecular mechanism of ALDH systematically.

(1) Gomez-Manzo et al. (2010) J Bacteriol 192, 5718-5724; (2) Miah R et al. (2021) Appl Microbial Biotechnol 105, 2341-2350.

−21−

(30)

-

N/C

N/C 0.08

- N/C 0.08

-

/NES GFP

NES-GFP Sci. Rep., 2016

NES-GFP NES-GFP

N/C

NES-GFP

CoQ

1 2 2 2 2 1,2,3,

1,2,3 1 , ² , ³

Q CoQ CoQ

CoQ10 0.2

php2 Php2 Php3 Php5 Php CCAAT

CoQ Php CoQ

php CoQ10

php2 php3 php5 CoQ

php2 php5 CoQ10

php3 CoQ10

php2 php3 php5 CoQ10 2.0

CoQ10

php

CoQ php2 php3 php5

CoQ PHB

−22 −

(31)

1 2

S- (SAM)

SAM S-

(SAH) SAH

SAH

mTORC1(mammalian target of rapamycin)

SAH mTORC1 SAH

Sch9 mTORC1

SAH

mTORC1 mTORC PP2A

SAH PP2A SAH mTORC

PP2A SAH

COP

1 1

COPII Sec12 Ino2

Ino4 Ino2/Ino4

COP

Ino2/Ino4

2 175 2 47

COP 175

8 BAP2 GGC1 FET3 TRF5 MSG5 YIP3 DSE4 YOR338W

COPII

−23 −

(32)

Tor-kinase complex 2 (TORC2)-Ypk1 TORC2-Ypk1

Orm1/2 Ypk1

Com2 YPK1 LCB1

Com2 YPK1 LCB1 -

( -Gal) LacZ YPK1 LCB1

Com2 Ptet-off-GFP-COM2

Com2 PYPK1-LacZ PLCB1-LacZ -Gal

Com2 5 2 Com2

6 Com2

Com2 YPK1 LCB1 YPK1 LCB1

sde

(6-Tsp)

BAR Pil1 6-Tsp pil1 6-tsp

SDS pil1 6-tsp SDS

6-Tsp

pil1 6-tsp SDS Suppressor mutants

for SDS sensitivity of eisosomal deletion mutant with temperature sensitivity (sde) 93

6 sde1 sde6 TOR2 LST8 AVO3

TORC2 pil1 6-tsp SDS

TORC2-Ypk1 pil1 6-tsp SDS

TORC2-Ypk1

sde TORC2-Ypk1

−24 −

(33)

Acidovorax cirulli

Aave_4606

Acidovorax cirulli

(Trx) (GSH)

Ralstonia solanacearm RipAY

Trx GSH A. citrulli

Aave_4606 RipAY RipAY

Aave_4606

Aave_4606 GSH

Aave_4606

Trx GSH Aave_4606

(aave_4606 ) (hrcV ) ,

GSH aave_4606 GSH

Aave_4606 RipAY Trx

GSH

Aurantiochytrium

1 1

Aurantiochytrium

8% Aurantiochytrium limacinum SR21

25% 27%

SR21 5

SR21

−25 −

(34)

EPA

sgRNA all-in-one vector

CO2 CO2

carbonic anhydrase2 (CA2)

Nannochloropsis oceanica NIES-2145 N. oceanica

carbonic anhydrase CA2 all-in-one vector

(pNAN1006) NIES-2145 CA2

f/2 CA2

CO2 0.15 0.15 3% 5% CO2 0.15 CO2

3%CO2

5%CO2

CA2 CO2

10

1-

-3-

Pattana Kakumyan¹ ²

3 3

1 2 3

1- -3-

1- -3- (KO )

1- -3-

Drosophila melanogaster

KO 1- -3-

Neoempheria dilatata

Aphelenchus avenae KO

1- -3-

−26 −

(35)

11

Eikenella corrodens

1 1

E. corrodens

E. corrodens 1073 luxS

luxS luxS

GalNAc GalNAc

luxS

12

Ralstonia solanacearum

Asmaa Ali Ahmed Ibrahim

MCP

Ralstonia solanacearum 22 8

14

R. solanacearum

PSD6

PSD6 8 8

mcp10 Mcp10

Mcp10 Mcp10

HPLC Mcp10

−27−

(36)

13

MCP

Pseudomonas protegens CHA0 mcp42

TLC 2

TLC HPLC

−28 −

(37)

N P S K

10% MGRL 2

P K

S N

N

vtc2 4 N

vtc2

N vtc2 N

vtc2

Nudix hydrolase

1 2

NAD(H) NADP(H)

Nudix hydrolase NADH AtNUDX6, 7 NADPH

AtNUDX19 /

Nudix hydrolase

AtNUDX6, 7 19

nudx7 nudx6/nudx7

nudx19, nudx6/nudx19

nudx6/nudx7 NADH, NADPH NADP(H)

nudx7 nudx6/nudx7 nudx6, nudx19

nudx6/nudx7 NAD(H), NADP(H)

−29 −

(38)

1 2

B2 ( : RF) FMN FAD

/ DNA

RF

purine permease (PUP)

PUP

PUP (AtPUP) AtPUP

20 AtPUP RF RF (rib5

) 2 AtPUP RF

2 AtPUP RF RF

12 RF 2 AtPUP RF

2 AtPUP

AtPUP-GFP

2 AtPUP

1 2

B2 RF: FMN

FAD /

FAD-responsive transcription factor 19 FRTF19 FRTF19

FRTF19 ES ES

FRTF19 RF RF

FRTF19 RF

RF FRTF19

FRTF19

FRTF19 FRTF19

− 30 −

(39)

1 1 2

1 2

Nicotiana tabacum

BAS RZS1 35S

PAP1 CHS

0.93 µg/gFW 4.5 µg/gFW

CPC

(Camellia sinensis var. sinensis)

R3 MYB CPC

CPC 6

CPC

CPC CPC

CPC

6 CPC

− 31−

(40)

MYB

(Arabidopsis thaliana)

MYB CAPRICE(CPC)

7

cpc MYB

MYB

(Col-0) cpc pCPC::CPC:GFP in cpc pCPC::TRY:GFP in cpc pCPC::ETC1:GFP in cpc pCPC::ETC2:GFP in cpc pCPC::ETC3:GFP in cpc

GFP

pCPC::TRY:GFP in cpc

GFP pCPC::CPC:GFP in cpc

pCPC::ETC1:GFP in cpc pCPC::ETC3:GFP in cpc

EGFR-ERK

1 1 2 1

1 2

Citrus sudachi SPE

SPE HaCaT BrdU

HaCaT SPE

EGF EGFR Raf-MEK-ERK

SPE EGF

EGFR Raf MEK1/2 ERK1/2 SPE

TNF- TNF- EGFR ERK1/2

SPE keratin1

keratin10 involucrin mRNA SPE

SPE mRNA

SPE EGFR-ERK

− 32 −

(41)

1 1 1

1) 2)

25 60 2 4

MeOH HPLC-DAD-MS UV

1) 2021

3C06-06 ²⁾ 2021 3C06-07

− 33 −

(42)

STZ

Tolulope Peter Saliu¹ ¹ ¹

1

streptozotocin STZ

STZ STZ

STZ

STZ 2

STZ 8 C57BL/6J 180 mg/kg STZ

4 STZ STZ 8 C57BL/6J

45% 4 50 mg/kg STZ 5

12 STZ+HFD

STZ DNA microarray

STZ apoptosis STZ+HFD

OGTT DPPIV AW WV

GLP-1

1 2 3

DPP IV

Sep-Pak C18 N5 10%

(N5-2 )(IC50=73.71 µg/ ml) N5-2 19.63% DPP IV

100.15% Ala-Va-Phe, Gly-Val-Phe, Glu-Val-Phe

Trp-Val ( 0.14% ; Trp-Val 15.24 mg/ 100 g N5-2 DPP IV IC50 ; 11.21 µg / ml )

5-2 Caco-2 400 mg/ml 20% DPP IV p<0.05

N5-2 DPP IV ICR (n= 8)

N5-2 (OGTT) N5-2 (100

mg/kg) 1 g/kg BW

30 p<0.05 60 N5-2

N5-2 DPP

IV K579 OGTT 0 30AUC 0

30AUC Trp-Val OGTT 100 mg, 300 mg, 1000 mg/kg

0 30AUC 0 30AUC 1000 mg 0 30AUC 21%,

0 30AUC 60 DPPIV

− 34 −

(43)

1 1 2 2

1 2

(G-Hes) G-Hes

8 C57BL 4 40 mg/kg

streptozotocin(STZ) 5 8 DN

DN STZ 1% G-Hes

8 G-Hes

Slc5a2 DN G-Hes

chemokine Ccl2

G-Hes NF- B DN

G-Hes G-Hes

B12

1 1

B12 (B12) B12 CoA

(Caenorhabditis

elegans) B12

B12

GMC101 25

1-42 B12 B12 B12

5 B12 B12

B12

− 35 −

(44)

8 37 45 LPS

1.0×10 cfu 3 1 µg/mL LPS

48 TNF- ELISA

TNF- Lactobacillus (Lb.) hilgardii H-50

Lb. homohiochii S-26 IL-1 IL-6 ELISA

LPS 4 Tnf- Il-1 Il-6 mRNA qPCR

LPS Lb. hilgardii H-50 Lb. homohiochii S-26 LPS

TNF- Lb. homohiochii S-26 Lb. hilgardii H-50

LPS TNF- IL-1 IL-6 Tnf- Il-1 Il-6 mRNA

45 Lb. hilgardii H-50

Lb. hilgardii H-50

Thanutchaporn Kumrungsee

1 1

1

Lactobacillus brevis 1059^T - GABA

fDREM fDREM

DREM TNF-

GABA

fDREM TNF- GABA

Balb/c 6-8 GABA-A + -

BCC 100 M 3 1% v/v DREM fDREM

0.2 mM GABA 1% v/v DREM 72

TNF- ELISA

BCC DREM TNF- fDREM

TNF- BCC GABA DREM

DREM GABA TNF-

fDREM GABA

− 36 −

(45)

RBL-2H3 -

WST-8 RBL-2H3

PI3K Akt

LCR

− 37−

(46)

-

− 38 −

(47)

2021 5 13 51

− 39 −

(48)

(49)

日本農芸化学会中四国支部第59回講演会主催：日本農芸化学会中四国支部

世話人：羽倉義雄

連絡先：739-8528 広島県東広島市鏡山1-4-4 広島大学大学院統合生命科学研究科ＴＥＬ：082-424-7938

E-mail ：[email protected]

(50)

支部からのお知らせ

1. 2021年度西日本・中四国・関西支部合同大会（第60回講演会）

開催日：2021年９月24日（金）〜 25日（土）

場所：かごしま県民交流センター，鹿児島大学郡元キャンパス内容：特別講演，シンポジウム，受賞講演，一般講演

一般講演申込締切：７月16日（金）

講演要旨提出締切：７月30日（金）

世話人：玉置尚德（鹿児島大学）

2. 支部創立20周年記念第61回講演会（例会）

開催日：2022年１月22日（土）

場所：高知大学物部キャンパス

内容：シンポジウム，受賞講演，一般講演世話人：島村智子（高知大学）

3. 支部創立20周年記念第38回市民フォーラム開催日：2021年８月21日（土）

場所：香川大学農学部

内容：招待講演，高校生によるポスター発表世話人：田淵光昭（香川大学）

4. 支部創立20周年記念第39回市民フォーラム開催日：2021年８月28日（土）

場所：島根大学大学ホール内容：招待講演

世話人：室田佳恵子（島根大学）

5. 支部創立20周年記念第32回若手研究者シンポジウム開催日：2021年７月30日（金）

場所：Web開催（愛媛大学）

世話人：米山香織（愛媛大学）

日本農芸化学会中四国支部事務局

〒739-8530 広島県東広島市鏡山 1-3-1 広島大学大学院統合生命科学研究科内

支部ホームページ：http://chushikoku.jsbba.or.jp/

E-mail： [email protected]

2021年（令和３年）6月12日発行

講 演 要 旨 集 - 日本農芸化学会中四国支部