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3-26 JAEA-Review 2010-065 Ion Beam Breeding of “Sake Yeast” and Test Brewing T. Masubuchi a) , O. Kamiyama a) , K. Satoh b) , Y. Hase b) and I. Narumi b) a) Gunma Industrial Technology Center, b) Radiation-Applied Biology Division, QuBS, JAEA We are trying to obtain high ethyl caproate producing Sake yeast by carbon ion-beam irradiation. Freeze-dried yeast was irradiated with carbon ion beams (220 MeV) in a range of dose from 50 to 300 Gy by AVF cyclotron at JAEA. We isolated 887 cerulenin resistant mutants of sake yeast “Kyokai-901”, “G2” and “No.227 (Ion beam mutant strain)”. Sake was brewed on 57 kg-scale Test Brewing with 2 mutants that had been isolated by last year’s irradiation experiments. No.227 seemed to hold enormous promise as a new type of yeast for quality sake because of its sweetness and uniqueness in flavor. 群馬県では現在オリジナルの吟醸用清酒酵母として「群馬KAZE酵母」を既に実用化しているが、消費者の嗜好の変化に対応した新たな酵母の開発をすべく、平成20年度よりイオンビーム照射による変異処理を行い、優良株の選抜を試みている。平成21年度は、更に優良な変異株を得るべく選抜を継続すると共に、それまでに得られた優良株の試験醸造を行い、醸造適性の確認を行った。選抜の指標は従来同様に吟醸酒の主要な香気成分であるカプロン酸エチルの生成能とした。【変異株の選抜】イオンビーム照射に供する試料の調製、照射後の処理は前報に準じて行った 1) 。照射は、TIARAのAVFサイクロトロンを用いて加速した炭素イオン(220 MeV 12 C 5+ )を50~300 Gy照射した。本年度は従来用いたきょうかい901号の他にG2酵母(群馬県保存株)、 No.227酵母(平成20年度にイオンビーム照射で得られた優良株)を親株として用いた。カプロン酸エチル高生成株を取得するための選択培地は、市川ら 2) の方法に準じセルレニン12.5~37.5 μM を含むYPD寒天培地を用いた。これに生育したセルレニン耐性変異株を齋藤ら 3) の方法を改変した発酵試験に供し、15 °Cで7日間の培養後に重量減少量(CO 2放出量)を測定し発酵力の目安とした。また、得られた上清をヘッドスペースガスクロマトグラフにより、イソアミルアルコール、酢酸エチル、酢酸イソアミル、カプロン酸エチルの各成分を定量して香気生成能を評価した。発酵試験は887株について実施したが、G2酵母からは優良株を得ることはできなかった。良好な香気生成能を示す変異株は主として100~150 Gyの照射区分で得られた。発酵試験で良好な17株について、布宮ら 4) の変即二段仕込みを改変した方法で小仕込み試験(総米200 g)を行った。いずれも対照とした群馬KAZE酵母(2号)よりも良好な香気生成能を示したため、発酵能などを考慮した上で次年度の試験醸造に供したい。【試験醸造】平成20年度に得られた変異株のうち、香気生成能が良好だったNo.231およびNo.227酵母を試験醸造に供した。対照として群馬KAZE酵母(2号)を用いた。試験醸造は群馬産業技術センター保有の醸造設備を用い、 40%精白山田錦を原料とした総米57 kgの吟醸造りを行った。 - 82 - No.231酵母は、醪の発酵状態を示す指標であるBMD 値の経時曲線が群馬KAZE酵母とあまり変わらなかったが、No.227酵母は醪後半のキレが緩やかであり、群馬KAZE酵母より発酵力は弱い(Fig. 1)。一方、 No.227酵母の醪中のカプロン酸エチル量は三者の中で最も多い(Fig. 2)。上槽した清酒を官能評価すると、 No.227酵母は甘さ、ふくらみなどの面で新しい香り傾向を示し、新たな吟醸酵母として期待が持てる。 Fig. 1 BMD in sake-moromi by Test brewing. Fig. 2 Ethyl caproate in sake-moromi by Test brewing. References 1) T. Masubuchi et al., JAEA Takasaki Ann. Rep. 2008 (2009) 84. 2) 市川英治 et al., 1988年度日本農芸化学会大会講演要旨集、435. 3) 齋藤久一 et al., 醸協 87 (1992) 915. 4) 布宮雅昭 et al., 醸協 90 (1995) 217.
3-27 The Effect of γ-Sterilization of Carrier Materials on the Shelf Life of Biofertilizer K. Tejima a) , T. Yokoyama b) , K. Satoh a) and I. Narumi a) a) Radiation-Applied Biology Division, QuBS, JAEA, b) Institute of Agriculture, Tokyo University of Agriculture and Technology Numerous studies on the inoculation technique of beneficial microorganisms to plants at agricultural fields have been carried out worldwide. However, except in a case of a low competition with native microbes, the practical technique that shows remarkable effect on the plant growth has not been established. Biofertilizer is a substance that holds beneficial microorganisms for plant growth (e.g. Rhizobia) in the carrier such as peat, perlite, charcoal and soil 1) . To use biofertilizer more efficiently, it is necessary to keep the amount of viable inoculants in the biofertilizer for a definite period of time. The inoculants survival in the biofertilizer could be affected by the physical and chemical properties of carrier materials and by the competition with native microbes in the carrier. Gamma-irradiation is expected to sterilize the carrier materials without changes in physical and chemical properties. In an effort to demonstrate the advantage of γ-sterilization, the survivals of inoculants were monitored to assess the shelf life of biofertilizer. Two kinds of peat (commercial sterilized peat moss from Canada and Japanese peat soil: Named “P1” and “P2”, respectively), Japanese forest subsoil of the Kanto Loam Formation (Named “L”), and three kind of Japanese cultivated soil (topsoil of the Andosol, Gray Paddy soils and Brown Fluvic soils: Named “S1”, “S2” and “S3”, respectively) were prepared. Except P1, the soil samples were air-dried and passed through a 2 mm sieve. All soil samples were stored at room temperature in the dark. In order to optimize the γ-ray dose to sterilize soils for the purpose of use as a biofertilizer’s carrier material, the survival rate of native microbes in the prepared soils were determined. P2, L, S1 and S2 were packed in plastic bags and irradiated with 1 k, 5 k, 10 k, 20 k, 30 k and 50 kGy of γ-rays. As a result, 50 kGy was determined to be suitable to sterilize soil (Fig. 1). This result was confirmed by additional experiments where P1 and S3 were also sterilized completely by 50 kGy γ-irradiation. As carrier materials, P1, P2, S1 and S3 were chosen. To make the soil-based artificial carriers, the S1 and S3 were mixed with charcoal powder (3:1) (Named “CS1” and “CS3”, respectively). Following the addition of 20 mL of sterilized water per 100 g of the carriers (P1, P2, CS1 and CS3), they were sterilized by 50 kGy γ-irradiation or autoclaving at 121 °C for 40 min. After that, water suspension (20 mL) of Bradyrhizobium japonicum strain USDA110 was inoculated to sterilized or non-sterilized carriers. The final density of inoculants in the biofertilizer was 6.6 × 10 7 cells g –1 . The biofertilizer was packed in plastic bags and stored for one month in the 4 °C or 30 °C. JAEA-Review 2010-065 - 83 - Before and after storage, viable inoculants in the biofertilizer were enumerated by the dilution plate method. Relative ratios of viable inoculants in the biofertilizer stored for one month are shown in Table 1. The relative ratio at pre-storage is defined as 1. As a result of storage at 30 °C, the relative ratio increased in the biofertilizer made from γ-sterilized or autoclave-sterilized carrier, though it tended to decrease in the biofertilizer made from non-sterilized carrier. The γ-sterilization was superior to the autoclave-sterilization in enhancing the inoculants survival. For P1, the relative ratio was high irrespective of sterilization treatment in this study because P1 had already been sterilized during manufacturing process. On the other hand, only a small change in the relative ratio was detected in the biofertilizer stored at 4 °C. Reference 1) K. Senoo and I. Narumi, FNCA Biofertilizer Manual (2006) 41. Fig. 1 Survival rates of native microbes in the different kinds of soils after γ-irradiation. Survival microbes were not detected at 50 kGy irradiation. Table 1 Relative ratios of the viable inoculants in the biofertilizer.
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JAEA-Review 2010-065 JAEA Takasaki
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JAEA-Review 2010-065 PREFACE This r
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JAEA-Review 2010-065 and pulsed hea
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JAEA-Review 2010-065 1-23 Studies o
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JAEA-Review 2010-065 3-24 Lethal Ef
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JAEA-Review 2010-065 4-07 Fabricati
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JAEA-Review 2010-065 5. Status of I
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JAEA-Review 2010-065 1-13 Alpha-rad
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1-01 Radiation Resistance of InGaP/
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1-03 Evaluation of Soft Error Rates
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1-05 Heavy-ion Induced Current in S
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Total Ionizing Dose Tolerance of Si
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1-09 Evaluation of Single Event Eff
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Hydrogen Diffusion in a-Si:H Thin F
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1-13 Alpha-radiolysis of Organic Ex
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Study on Stability of Cs·Sr Solven
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Irradiation Effect of Gamma-Rays on
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1-19 Development of Radiation Resis
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1-21 Alpha-Ray Irradiation Damage o
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1-23 Studies on Microstructure and
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1-25 Effect of Groundwater Radiolys
Page 47 and 48: 1-27 Simulation of Neutron Damage M
Page 49 and 50: 1-29 Irradiation Hardening in Ion-i
Page 51 and 52: 1-31 Preparation of Anion-Exchange
Page 53 and 54: 1-33 Radiation-Induced Graft Polyme
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Page 58 and 59: 2-02 Development of Zwitterionic Mo
Page 60 and 61: Modification of Hydroxypropyl Cellu
Page 62 and 63: The Recovery of Precious Metals Usi
Page 64 and 65: 2-08 ESR Study on Carboxymethyl Chi
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Page 69 and 70: JAEA-Review 2010-065 3-28 Electron
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Page 74 and 75: 3-02 Mutagenic effects of He ion pa
Page 76 and 77: 3-04 Functional Analysis of Flavono
Page 78 and 79: 3-06 Generation New Ornamental Plan
Page 80 and 81: 3-08 Stability of Flower-colour Mut
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Page 84 and 85: 3-12 Producing New Gene Resources i
Page 86 and 87: 3-14 Production of Soybean Mutants
Page 88 and 89: Assessment of Irradiation Treatment
Page 90 and 91: 3-18 Effect of Different LET Radiat
Page 92 and 93: 3-20 JAEA-Review 2010-065 Fungicide
Page 94 and 95: 3-22 JAEA-Review 2010-065 FACS-base
Page 96 and 97: 3-24 Lethal Effects of Different LE
Page 100 and 101: 3-28 Electron Spin Relaxation Behav
Page 102 and 103: 3-30 Target Irradiation of Individu
Page 104 and 105: 3-32 Carbon-ion Microbeam Induces B
Page 106 and 107: 3-34 Biological Effects of Carbon I
Page 108 and 109: 3-36 Difference in Bystander Lethal
Page 110 and 111: 3-38 Analysis of Lethal Effect Medi
Page 112 and 113: 3-40 Irradiation with Carbon Ion Be
Page 114 and 115: 3-42 Expression of Two Gelsolins in
Page 116 and 117: 3-44 Analysis of Bystander Cell Sig
Page 118 and 119: 3-46 Quantitative Evaluation of Ric
Page 120 and 121: 3-48 Visualization of 107 Cd Accumu
Page 122 and 123: 3-50 Uniformity Measurement of Newl
Page 124 and 125: 3-52 Imaging and Biodistribution of
Page 126 and 127: 3-54 Improvement of Spatial Resolut
Page 128 and 129: 3-56 The Optimum Conditions in the
Page 130 and 131: 3-58 Evaluation of Cisplatin Concen
Page 132 and 133: 3-60 JAEA-Review 2010-065 Analysis
Page 134 and 135: 3-62 JAEA-Review 2010-065 Sensitivi
Page 136 and 137: JAEA-Review 2010-065 4-14 The Effec
Page 138 and 139: JAEA-Review 2010-065 4-39 Relations
Page 141 and 142: 4-01 Hydrogen Gasochromism of WO3 F
Page 143 and 144: 4-03 Synthesis of Single-Crystallin
Page 145 and 146: 4-05 Synergy Effects in Electron/Io
Page 147 and 148: Fabrication of Diluted Magnetic Sem
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4-09 Gas Permeation Characteristics
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4-11 Control of Radial Size of Poly
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4-13 Behavior of N Atoms in Nitridi
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4-15 JAEA-Review 2010-065 Annealing
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Low Temperature Ion Channeling of F
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4-19 Radiation-Induced Electrical D
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4-21 Study on Cu Precipitation in E
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4-23 Evaluation of Fluorescence Mat
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4-25 Evaluation of the ZrC Layer fo
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4-27 Structure Analysis of K/Si(111
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4-29 LET Effect on the Radiation In
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4-31 Stabilization of Measurement S
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Systematic Measurement of Neutron a
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4-35 Measurement of Neutron Fluence
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4-37 Evaluation of the Response Cha
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4-39 Relationship between Internucl
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4-41 Analysis of Radiation Damage a
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4-43 Positive Secondary Ion Emissio
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4-45 JAEA-Review 2010-065 Ion Induc
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4-47 Fabrication of Dielectrophoret
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4-49 Fast Single-Ion Hit System for
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4-51 Development of Beam Generation
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JAEA-Review 2010-065 5. Status of I
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Operation of the AVF Cyclotron T. N
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Operation of Electron Accelerator a
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5-06 FACILITY USE PROGRAM in Takasa
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5-08 Radioactive Waste Management i
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Appendix 2 List of Related Patents
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Appendix 4 Type of Research Collabo
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表1.SI 基本単位基本量 SI
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