本研究以幾丁聚醣作為固定化擔體，利用物理性膠凝作用，針對惡臭假單胞菌進行包埋固定化，作為一代謝酚的生物觸媒，此一固定化擔體有兩項性質是本實驗所關心的議題：
1. 顆粒表層與剖面形態：利用電子顯微鏡觀察不同膠凝條件下形成的顆粒，尋求表層孔洞分布程度高且孔徑小於0.2μm，避免菌體逸漏，達成固定化目的，而顆粒的核心層勿過於緻密，方可提供菌體足夠的生長空間。
2. 顆粒內包埋的活菌數：已固化成型的幾丁聚醣顆粒不易再度溶解，菌體的定量更加困難，採間接探討包埋過程中流失的菌數，與經包埋溶液接觸後的失活情形，來反推顆粒內包埋的活菌量。

利用2%幾丁聚醣溶液在三聚磷酸鈉中所膠凝的顆粒，可耐受曝氣測試與震盪測試，可見幾丁聚醣顆粒強度的穩定性佳。該顆粒表面孔徑為1~6μm，應該會有菌體逸漏的問題。幾丁聚醣分子量愈大與去乙醯程度愈高，都有助於幾丁聚醣滴入膠凝劑時，可以膠凝成顆粒狀。膠凝顆粒隨幾丁聚醣濃度增加，其顆粒核心愈密實，將會對菌體的生長空間有所限制。菌體一經幾丁聚醣溶液接觸後，立即失活，可能是幾丁聚醣的抑菌性與偏酸的幾丁聚醣溶液所致。

In this study, chitosan is used as an immobilization matrix. Pseudomonas putida is entrapped by physical iontropic gelation with chitosan and the resulting beads are used as a biocatalyst for the biodegradation of phenol. Two important issues were investigated about these beads:
1. The surface and cross-section morphology of the beads: the structure of the beads formed by different iontropic gelation procedures were observed by scanning electron microscopy (SEM). To avoid bacteria leakage and attain an effective immobilization, the high surface porosity and small pore diameter that is less than 0.2μm are sought. In addition, the core of the beads shouldn’t be too dense, so as to provide enough growing space for bacteria.
2. The viable bacteria concentration inside the bead: the quantification of bacteria is difficult since the lysis for the chitosan bead was hard. Thus, the following experiments were performed. One is the concentration of the bacterial loss during entrapment. The other is the viable ratio of the bacteria after contact with the solution used for immobilization. Henceforth, the viable bacteria concentration entrapped inside the bead can be calculated.

Chitosan beads formed by using 2% chitosan solution gelation in 1% sodium tripolyphosphate solution is strong enough to bear the aeration and shaking tests. This indicates the stability of the chitosan beads were satisfactory. The surface pore diameter of the beads formed is ranging from 1 to 6 μm. This might result in the leakage of bacteria entrapped. High molecular weight and high degree of deacetylation of the chitosan can lead to a better bead formation in the gelation solution. In addition, the core of the beads will become denser with the increase of the chitosan concentration, and this will limit bacteria growth. Most of all, it was found that the bacteria is denatured after its contact with chitosan solution. This may be attributed to the antibacterial property associated with chitosan as well as the acidic environment of the chitosan solution.

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