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本文報告了一種簡便高效的細菌磨,其主要構成部分包括一段横置的磨沙硬質滾筒和凹面瓷座。用5,000×g轉速,20分鐘離心收集的大腸桿菌漿作試驗。這種菌漿含水量約為乾菌重的5倍。加入1.3—1.5倍重的玻璃粉(顆粒小於3μ),混合均匀,研磨20—30秒,細菌裂碎率保證在99.99％以上。 將研磨所得混合物懸浮於磷酸鹽緩衝劑中,離心分離得到全提取液。全提取液再以20,000×g轉速,2小時離心分離,將酶劑分為澄清提取液和從碎菌沉澱製成的碎菌懸浮液兩部分。後者所殘留的活菌的氮約佔全氮的10~(-3)—10~(-4)％。利用同一批培養出來的細菌各次研磨製得的酶劑,其酶活力的可重複性程度很高,以琥珀酸氧化酶系的活力來說,各次平均離差範圍僅±10％。不需要加任何輔酶輔基,這兩部分酶劑都能够直接利用空氣中的氧來氧化琥珀酸、L-蘋果酸、L-乳酸、甲酸。澄清提取液部分還可以氧化L-谷氨酸、L-門冬氨酸、延胡索酸和乙醇,並且當它和碎菌部分酶劑混合時,除琥珀酸、甲酸和L-乳酸外對其他底質的氧化活力都大為提高。
This article reports a simple and efficient bacterial mill, the main components of which include a horizontal section of sandblasted hard drum and concave porcelain seat. The E. coli slurry collected by centrifugation at 5,000 x g for 20 minutes was tested. This mycelial water content is about 5 times the weight of dry bacteria. Add 1.3-1.5 times the weight of glass powder (particles less than 3μ), mixed evenly, grinding 20-30 seconds, the bacterial fragmentation rate of 99.99% guaranteed. The resulting mixture was suspended in phosphate buffer and centrifuged to obtain a total extract. The whole extract was centrifuged again at 20,000 × g for 2 hours to separate the enzyme into clarified extracts and a suspension of the bacterial cells that had been obtained from the sedimentation of the broken bacteria. The latter remaining viable bacteria nitrogen accounted for about 10 ~ (-3) -10 ~ (-4)% of total nitrogen. Using the same batch of bacteria produced by grinding each enzyme preparation, the enzyme activity of highly reproducible degree of succinic acid oxidase system activity, the average deviation range of only ± 10%. Do not need to add any coenzyme prosthetic groups, these two enzyme agents are able to directly use the oxygen in the air to oxidize succinic acid, L-malic acid, L-lactic acid, formic acid. Clarification extract part can also be oxidized L-glutamic acid, L-aspartic acid, fumaric acid and ethanol, and when it is mixed with the broken part of the enzyme, in addition to succinic acid, formic acid and L- The oxidation activity is greatly improved.