臺灣博碩士論文加值系統

隨著人類生活品質的提升，大家也開始注重與大自然的相處，除了溫室氣體排放外，由於工業區廢水排放常見重金屬含量偏高，尤其六價鉻水體污染為當今嚴重的環境問題，而含有鉻的產品若沒經過妥善回收處理，鉻汙染將遍及土壤及水中，對臺灣的環境造成嚴重傷害，其中Cr(III)是人體必需營養素，可幫助人體使用糖分、蛋白質及脂肪的必須營養素，且三價鉻造成的毒性影響也較六價鉻低許多，而高濃度的六價鉻會傷害鼻子及造成癌症，食入後也會造成貧血或腸胃的損傷。因此，利用Chlorella具有吸附廢水中重金屬的能力，探討微藻吸附Cr(VI)的效果。綜合實驗結果，已知微藻移除重金屬速率約在2.50 ~ 3.33 mg /L．day，進而延伸探討微藻如何移除Cr(VI)，發現綠藻內含有具抗氧化功能的谷胱甘肽(GSH)，在微藻進行重金屬移除的過程中，谷胱甘肽(GSH)也可能少部分的使Cr(VI)轉變為Cr(III)，且當微藻中谷胱甘肽(GSH)濃度越大，將可以還原越多的Cr(VI)，但反應順序為微藻還原酵素皆反應完後，才會由抗氧化劑谷胱甘肽(GSH)反應，證實了微藻在移除Cr(VI)的過程中，不只是將Cr(VI)吸附還有少部分的Cr(VI)被綠藻還原酵素與抗氧化劑GSH還原成Cr(III)。
此外，在台灣，許多生活廢水和生產污水時常在未經過處理就直接排入河川中，使得含有高營養物質、磷、有機物等廢水直接汙染了河川，導致河川生態受到影響和破壞，負荷不了這麼多汙染的河川，也逐漸的變得混濁和惡臭及蚊蟲孳生。因此，利用Chlorella具有代謝有機物等高營養物質的能力與培養週期短且穩定等特性，探討利用批次（Batch）及連續式（Continuous）反應培養C. vulgaris並同時去除廢水中化學需氧量(COD)。綜合實驗結果，在批次培養方面，以稀釋50 %的葡萄糖碳源發酵紅酵母後之廢水材料最適合作為微藻的生長環境，培養10天約可以降低廢水中7000 mg/L 的化學需氧量，在連續式培養上，以水力停留時間5天最適合微藻生長以及菌體累積，培養10天後，一天約可以降低廢水中10000 mg/L的化學需氧量，找出最佳的培養條件後，進而探討放大反應體積與改變反應形式對於降低廢水中化學需氧量之效果，得到在批次培養下，放大為5L反應器與改變反應形式，淨化效果提升至10天降低廢水中化學需氧量10000 mg/L，而連續式培養下，放大為5L反應器與改變反應形式，設定HRT 5天，淨化效果提升至10天後，每天降低廢水中化學需氧量 14000 mg/L，發現改變反應體積與反應形式可以提升降低廢水中化學需氧量的效果，也得到對於微藻降低水中化學需氧量以連續式反應為最佳的反應設置條件，整體系統可以維持微藻藻種濃度約2.0 g/L，且具有更好的化學需氧量移除效果與廢水使用量。
探討完重金屬的移除與降低廢水化學需氧量之外，最後討論利用串聯培養並進行固碳實驗，本次研究將黏紅酵母菌Rhodotorula glutinis及小球藻Chlorella vulgaris之生物反應器相互串聯，利用R. glutinis在發酵過程中代謝碳源並釋出CO2，再將CO2供給C. vulgaris做為碳源使用。由此系統供給CO2濃度培養C. vulgaris得到藻體平均濃度為1.51 g /L，故證實R. glutinis所釋出之CO2可用來培養C. vulgaris。此外，R. glutinis釋出之氣體，CO2含量大約為2.0 ％，每一克的紅酵母菌體，每天可產出0.142 kg CO2 ；而經由微藻固碳反應後，C. vulgaris釋出之CO2含量大約為0.2 ％，每一克的微藻藻體，每天可固定1.08 kg CO2生成綠藻細胞，故此串聯系統之微藻C. vulgaris可有效固定R. glutinis 所釋出之CO2達35.6 ％，結果顯示此一串聯系統可有效利用好氧系統排放之二氧化碳，達到系統淨排放及循環共生之目的。

With the improvement of human quality of life, environmental problems at the same time become the focus of global attention. Green house gas emission and heavy metals pollution has become a global issue of concern due to their higher toxicities. Especially, Cr(VI) is introduced in the environment mainly as a consequence of its industrial use and it has been causing serious environmental pollution due to its carcinogenicity.
In this study, the possible use of Chlorella vulgaris biomass as an alternative biosorbent for Cr (VI) removal was investigated. Therefore, the use of C. vulgaris has the ability to adsorb heavy metals in wastewater, and to explore the effect of microalgae adsorbing heavy metal Cr(VI). The results showed that microalgae can removal of Cr (VI). And then extend the discussion of microalgae how to remove heavy metals Cr (VI). It was found that glutathione (GSH) in C. vulgaris could decrease the heavy metal Cr(VI) into Cr(Ⅲ) in the process of heavy metal removal in microalgae, and the greater the concentration of glutathione (GSH), the greater the reduction of heavy metals Cr (VI). But the reaction sequence is microalgae reduction enzyme exhausted, the antioxidant glutathione will be the reaction. It is confirmed that Cr (VI) is not only adsorbed by algae, but also a small part of Cr (VI) is reduced to Cr(Ⅲ) by reduction enzyme and antioxidant during the removal of heavy metal Cr (VI) by microalgae.
Additionally In Taiwan, many domestic sewage and production wastewater are often discharged into rivers, untreated, high levels of nutrition, phosphorus, organic matter and other waste water directly pollute the river, causing the river ecology to be affected and damaged, The river suffered from severe pollutions has gradually become muddy and foul, and then lots of mosquitos and insects have bred in it. Therefore, use of Chlorella has the ability to metabolize organic matter and other high nutrient capacity and it's culture cycle is short and stable, to explore the use of batch and continuous reaction to culture C.vulgaris and remove chemical wastewater (COD). According to the comprehensive experimental results, 50% of glucose carbon source's R. glutinis fermentation wastewater is the most suitable microalgae growth environment. The chemical oxygen demand can be reduced about 10000 mg/L day by batch. In the continuous culture study, the 5-days hydraulic retention time is most suitable for microalgae growth and cell accumulation, culture for 10 days can reduce wastewater chemical oxygen demand 7000 mg/L in 10 days .The next discussion is best condition of scale up 5 times of culture's reaction volume and changes the reaction form. In the batch culture, the chemical oxygen demand in the 50% wastewater can be reduced to 10000 mg /L of COD in 10 days, and the chemical oxygen demand in the wastewater can be reduced by about 14000 mg/L in the HRT 5 days. It is found that the scale up reaction volume and change the reaction form can increase the degree of removing chemical oxygen demand in the wastewater, and the best reaction conditions for the reduction of chemical oxygen demand in the continuous reaction were obtained. The whole systom can maintain algal concentration of about 2.0 g/L, with good chemical oxygen demand removal effect.
Furthermore, in this sustainable cultivation system of aerobic yeast -Rhodotorula glutinis and photosynthetic microalgae -Chlorella vulgaris, a yeast and a microalgae were grown in two separate reactors connected by their gas transportation. The aerobic yeast provides CO2 for the growth of microalgae via photosynthesis process as both carrying out the production of lipids, and efficient CO2 fixation by Chlorella vulgaris. Moreover the aerobic yeast R. glutinis provides 0.142 kg CO2 / Day．g Biomass and C. vulgaris can utilize 1.08 kg CO2/ Day．g Biomass. Microalgae can utilize CO2 of emission gas from the yeast fermenter efficiently up to 35.6 %. It was demonstrated that this sustainable cultivation system of the yeast Rhodotorula glutinis and the autotrophic growth of the microalgae Chlorella vulgaris was successful in the reduction of CO2 emission.

中文摘要 I
Abstract III
第一章 緒論 1
第二章 文獻回顧 2
2.1微藻 2
2.1.1微藻簡介 2
2.1.2小球藻簡介 3
2.1.3微藻培養環境因子 4
2.1.4微藻培養方式 8
2.2環境中六價鉻之特性及汙染 9
2.2.1鉻物化特性 9
2.2.2環境中六價鉻汙染來源 9
2.2.3六價鉻之危害 10
2.2.4法令規定 10
2.3利用微藻移除重金屬 11
2.3.1微藻移除重金屬簡介 11
2.3.2微藻移除重金屬機制 13
2.4利用微藻移除廢水有機物 16
2.4.1 汙水處理概論 16
2.4.2微藻移除廢水簡介 18
2.4.3微藻淨化廢水機制與優點 19
2.5生物反應器串聯培養系統 21
2.5.1生物反應器串聯培養系統簡介 21
2.5.2綠藻固碳應用簡介 22
第三實 驗材料與方法 23
3.1 實驗材料 23
3.1.1 實驗藻種及廢水樣品 23
3.2實驗儀器 27
3.3分析方法 29
3.3.1光照強度測定方法 29
3.3.2藻體濃度測量方法 （吸光值測量藻體濃度） 29
3.3.3 葡萄糖濃度分析方法 29
3.3.4六價鉻濃度分析方法 30
3.3.5鉻價數轉換分析方法（XANES） 30
3.3.6化學需氧量(COD)測量方式 31
3.3.7谷胱甘肽(GSH)測量方式 31
3.3.8綠藻酵素測量方式 32
3.3.9 二氧化碳測量方式 32
3.4實驗方法 33
3.4.1藻種保存及活化 33
3.4.2菌種保存 33
3.4.3接菌 33
3.4.4培養基組成 34
3.4.4.1藻種培養基組成 34
3.5實驗架構 35
3.6實驗培養條件 36
3.6.1批次系統與連續式系統之利用紅酵母上清液對小球藻培養 36
3.6.1.1 500 ml C. vulgaris光合反應器批次發酵程序 36
3.6.1.3 5 LC. vulgaris批次系統與連續式系統光合反應器程序 37
3.6.2微藻C. vulgaris移除Cr(VI) 38
3.6.2.1微藻C. vulgaris還原酵素移除Cr(VI)培養程序 38
3.6.2.2微藻C. vulgaris谷胱甘肽(GSH)移除Cr(VI)培養程序(時間) 39
3.6.2.3微藻C. vulgaris谷胱甘肽(GSH)移除Cr(VI)培養程序(濃度) 39
3.6.3循環共生系統黏紅酵母菌與小球藻之串聯培養 40
3.6.3.1串聯50 L R. glutinis發酵槽及20 L C. vulgaris光合反應器重複饋料批次（Repeated Fed-batch）發酵程序 40
3.7實驗裝置圖 42
3.7.1微藻C. vulgaris移除Cr(VI) 42
3.7.2微藻批次反應培養 43
3.7.3微藻連續式反應培養 44
3.7.4連續式及批次5 L 放大反應與改變反應形式培養 45
3.7.5 20.0氣升式反應器與50 L黏紅酵母發酵槽反應器之串聯設備 46
第四章 結果與討論 48
4.1小球藻移除Cr(VI)探討 48
4.1.1探討小球藻C. vulgaris在利用綠藻酵素下進行生物機制移除Cr(VI)。 49
4.1.2探討小球藻C. vulgaris在利用綠藻藻體內谷胱甘肽(GSH)下進行非生物機制移除Cr(VI)。 51
4.2微藻C. vulgaris降低廢水中化學需氧量(COD) 57
4.2.1測試將綠藻培養於含有粗乾油廢水之實驗效果 58
4.2.2測試將綠藻培養於不含粗乾油廢水之實驗效果 60
4.2.3利用批式反應培養綠藻並去除廢水中化學需氧量探討 62
4.2.4利用連續式反應培養綠藻並去除廢水中化學需氧量探討 65
4.3串聯培養實驗 70
4.3.1循環共生系統黏紅酵母菌與小球藻之串聯培養 70
4.3.2串聯50 L R. glutinis發酵槽及20 L C. vulgaris光合反應器重複饋料批次（Repeated Fed-batch）發酵程序 70
第五章 結論與未來展望 73
5.1結論 73
5.2未來展望 75
參考文獻 76
附錄 81
作者簡歷 85

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