Yasuyuki Kaneko

Treatment of phenolic wastes byAureobasidium pullulans adhered to the fibrous supports

SummaryBiological treatment of waste water containing a large amount of phenol was carried out by using a phenolassimilating fungus,Aureobasidium pullulans No. 14 adhered (“semi-immobilized”) to fibrous asbestos. The column reactor employed for oxidative degradation of phenol consisted of a cylindrical glass column containing plastic nets.During 27 days operation, it was observed that: 1) The phenol removal capacity of the reactor gradually increased during the first 10 days, reaching a stable level. 2) The best phenol removal capacity (50 mg phenol removed/h/ liter of reactor volume) was obtained when an artificial waste water containing up to 1,200 μg/ml phenol was applied to the reactor. 3) Much higher concentrations of phenol (e.g. 1,700 μg/ml) brought about a marked decrease in the phenol removal capacity (40–50 mg/h/liter). 4) Satisfactorily stable operation was achieved using the semiimmobilized mycelia ofAureobasidium pullulans, whose active state could be checked by observing the thick, black-colored biomass which is characteristic of the genusAureobasidium and covered the plastic nets inside the glass column reactor.

Serial production of nucleoside-5′-triphosphates and uracil from 3′-mononucleotides by intact yeast cells

SummaryNucleoside-5′-triphosphates such as 5′-ATP and 5′-GTP can be produced efficiently and continuously from 3′-mononucleotides such as 3′-AMP and 3′-GMP by a series of processes consisting of two reaction phases using dried cells of Candida sp. N-25-2 (a hydrocarbon assimilating yeast). Moreover, incidentally to the 5′-triphosphates, free uracil is yielded almost stoichiometrically from 3′-CMP and 3′-UMP which, as is well known, are main concomitant products depolymerized from RNA. Uracil is then also available for many usage.

Studies on the Production of Hyaluronidase by Actinomycetes Part II

Streptomyces hyalurolyticusがhyaluronidase (HA-ase)を生産するための最適培地を検討した.HAaseはこれらの培地により構成酵素として産生され,生育と酵素生産とは並行しており,ジャーファーメンター培養では40 hrで最高46 TRU/mlに達した. 培養液よりDEAE, CMセルローズカラムクロマトグラフィー,有機溶媒沈澱,硫安分画等の常法によりHA-aseを精製し,33, 100 TRU/mg蛋白の精製標品を得た.この標品によりHA溶液は著しく粘度を降下させる. Ag+,Hg2+,pb2+等の金属イオン,I2,KMnO4等の酸化剤,モノヨード酢酸,N-bromosuccinimide, ρ-hydro-xymercuribenzoate等のSH試薬で阻害をほぼ完全にうけるが,これらの点は睾丸や細菌類のHAaseと性質を異にする.ヘパリン,コンドロイチン硫酸,ポリビニールアルコール硫酸等の高分子硫酸エステルによってもこの酵素は著しくHA分解作用を抑えられる.他の起源のHAaseと異なり,本酵素は精製を充分進めても決して不安定にはならない

Studies on the Metabolism of Polynuclear Aromatic Hydrocarbons by Microorganisms Part II

(1) 各地で採取した試料332個より,集積培養によりフェナンスレン資化菌22株を分離した. (2) 22株のフェナンスレン代謝産物を薄層クロマトグラフィーで比較し,すべてが1-hydroxy-2-naph-thoic acidを生成するのを認めた. (3) 分離菌のうち生育の良好なS-210, 592株の菌学的性質を検討し,いずれもPseitidomonas aeruginosaと同定した. (4) S-210, 592株はフェナンスレン,ナフタレンに資化能を示したが,アンソラセン,ベンツアンソラセンの資化能を示さなかった.

Studies on the Nucleases Produced by Microorganisms:Part III. Properties of Partially Purified Phosphodiesterase (5'-Nucleotide-Forming) Produced by Pellicularia sp

The properties of crude phosphodiesterase forming 5′-mononucleotide of Pellicularia H-II were investigated on its metal requirement, pH response for activity and so on. The dialyzate of crude PDase against distilled water became partly inactive, but was recovered with Zn++, Mn++ and Mg++, whereas completely inactivated dialyzate against EDTA was restored specifically with only Zn++The optimum pH of PDase activity was 5.0 and that of ribonuclease 4.0. The crude PDase was partially purified by acetone fractionation and Amberlite IRC-50 (XE-64) or CM-cellulose column chromatography. Two PDase and a RNase activities were recognized.Pellicularia PDase was found to be of new type according to its Zn++ dependency and non-activity towards bis-p-nitrophenyl phosphate.

Studies on the Glutamic Acid Fermentation. Part I

愛知県下の汚水からグラム陽性,無胞子,非運動性の好気性桿菌で糖から多量のL-グルタミン酸を生成するものを分離した.この菌は10%グルコース培地において振盪培養により4日間で対糖40%(培地1ml中40mg)以上のL-グルタミン酸を蓄積する.またその細菌学的性質から従来知られていない新菌と認め, Microbacterium salicinovorum nov. sp.と命名した.

Proteolytic Enzymes of Butyl Bacteria. Part III

アセトン・ブタノール菌,東大314号菌の生産する蛋白分解酵素と2個金属イオンとの関係につて試験して次の結果を得た. (1) 10-4MのEDTAによって殆んど完全に失活する.この失活は等モル量のCo++で完全に回復する. (2) EDTAと長時間,比較的高温に放置するとCo++を添加しても回復は小さくなるが,低温の場合には殆んど可逆的に活性は発現する. (3) EDTAに対し低温透析処理を行った結果,この失活はCo++, Mn++, Zn++, Fe++等で回復した.特にCo++及びMn++は有効である. (4)酵素液を放置する場合の失活に対してはMn++が保護作用を示し, EDTA処理酵素の失活防止には, Co++, Mn++, Zn++, Ca++等が有効である. (5)本実験より, EDTAによる失活は可逆的であり, EDTA処理酵素の変性(失活)は不可逆的に進行すると考えられる.