本研究所採用的菌株Acinetobacter radioresistens為革蘭氏陰性桿菌，篩選自廢水汙泥之嗜鹼性脂肪酵素生產菌，是一野生菌，較不易釋放出細胞內部的物質；最適合的培養溫度為30℃，所生產的脂肪酵素分子量為38 kDa、等電點為4.5，此酵素在pH 7.0~10.0及溫度30℃以下的室溫中具有良好的穩定性；有清潔劑產品工業的應用價值。
本脂肪酵素會分佈在水相及油水界面，添加1.5% 乙二醇可促進油水界面上酵素之回收。硫酸銨沉澱劑是實驗中較佳的選擇，回收率可達90%。提高起始蛋白質濃度與添加Na+、K+、Ca2+及Mg2+，實驗可得更好的回收效果。
本文提出新的策略之一：以水-正十六烷之油水界面行疏水性吸附，自醱酵液中回收脂肪酵素。利用此界面吸附之特性，產生大量的水-正十六烷界面，脂肪酵素可扮演乳化劑的角色而吸附在界面上，形成可與水分開的乳化相。藉著離心，水和正十六烷的乳化相可輕易分開。在此回收程序中，正十六烷並未被消耗，可回收再使用。此吸附遵循Langmuir吸附平衡關係，能以模擬預測吸附量和酵素活性。連續式多階段重複操作，結果濃縮倍數有4.2倍，回收率達67%。添加10%硫酸銨能提升濃縮倍數到4.4，回收率至79.8%。
本文提出新的策略之二：表面吸附液相正十六烷的不織布，可當成一吸附劑，以回收脂肪酵素。此酵素吸附於液態正十六烷表面上的量比固態正十六烷強很多；正十六烷的熔點介於16-18℃之間，利用小幅溫度變化，於液態正十六烷做吸附，而在固態正十六烷做脫附。在批次操作由於平衡的限制，所以濃縮倍數較低，在管柱實驗則可增加吸附劑的量，以提高濃縮倍數。吸附與脫附行為亦遵循Langmuir吸附平衡關係，利用此式在批次與管柱研究中得到良好的預測值，成功描述了吸附與脫附現象。
本文提出新的策略之三：在兩相流動超過濾中，以正十六烷取代空氣，減少因溶質產生的濃度極化現象。利用正十六烷的黏滯係數比空氣高，於兩相流動時可產生更大剪應力。以正十六烷/液的超過濾方式，可提昇脂肪酵素的濾出液通量與回收率，且沒有酵素失活的問題。以另外的系統，牛血清蛋白和糊精檢測濾出液通量，結果在通正十六烷於超過濾系統中，促進果效與脂肪酵素類似，而且都比空氣佳。若使用肌酸酵素溶液，通正十六烷後的濾速與酵素回收率也有明顯提昇。

The strain used in this study was A. radioresistens, a Gram negative bacterium isolated from wastewater sludge. The lipase produced is extracellular and has a molecular weight of 38,000 (Ng, 1998). The optimum temperature and optimum pH of this enzyme are 37℃ and 10.0, respectively. The lipase produced was found to distribute between the aqueous phase and the oil- water interface. The fraction of lipase in the aqueous phase could be increased to 85% if 1.5% (v/v) of ethylene glycol was added to the broth; nevertheless, further addition of ethylene glycol led to decreases in the lipase recovery For A. radioresistens lipase, the enzyme stabilizer could be Na+, K+, Ca+2 or Mg+2 at a concentration of 10 mM.
A. radioresistens lipase has a strong tendency to be adsorbed on the n-hexadecane-water interface. Taking advantage of this feature, we developed a recovery process by generating a lot of n-hexadecane-water interface in the enzyme solution. Lipase, acting as the emulsifier, could then be adsorbed at the interface, and form a separate emulsion phase. By centrifugation, the harvested emulsion phase again split into n-hexadecane and aqueous solution; thus concentrating the lipase. In this process, n-hexadecane was not consumed and could be reused. The adsorption process followed the Langmuir isotherm, and a multiple-run operation was recommended to enhance the enzyme recovery. The present process might be generally applicable to lipase recovery.
A. radioresistens lipase has a much higher affinity for liquid n-hexadecane than for solid n-hexadecane. Taking advantage of this feature, we immobilized n-hexadecane on a hydrophobic nonwoven fabric as the adsorbent to recover the lipase from fermentation broth. Above the melting point of n-hexadecane, we performed the adsorption, and below the melting point, the adsorbed lipase was then desorbed. There are no extraneous materials needed for desorption, the range of temperature shift can be quite small, and the enzyme will not denaturalize. The adsorption isotherm of lipase on the adsorbent follows the Langmuir model. Using the isotherm, we were able to simulate both batch and column adsorption/desorption processes. In batch process, the concentration factor is restrained by equilibrium; however, in column process, the concentration factor is proportional to the amount of adsorbent used.
n-Hexadecane has a much higher viscosity than air and hence can generate a much higher shear force in a two-phase flow process. Taking advantage of this feature, we mixed protein solutions with n-hexadecane in the operation of a crossflow ultrafiltration, and found that this was more effective in reducing concentration polarization than that with air sparging. For ultrafiltration of a crude lipase solution, both of permeate flux and activity recovery were significantly enhanced. In addition, there was no evidence of enzyme inactivation owing to the use of the two-phase flow. The n-hexadecane/water two-phase flow could have a general applicability in ultrafiltration practices.

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