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高耐鎘菌株P(guān)DA-H的篩選及活性研究 摘 要 當(dāng)下，土壤重金屬污染不斷加重，在我國，土壤鎘（下文簡稱Cd）污染尤為顯著，造成了許多危害。重金屬污染土壤的微生物修復(fù)作為一種低成本、操作方便、應(yīng)用前景廣泛的綠色修復(fù)技術(shù)，倍受國內(nèi)外研究者關(guān)注。本文以從蘇州地區(qū)典型污染場地土壤與高有機(jī)質(zhì)土壤中篩選出的耐Cd菌株P(guān)DA-H為研究對象，進(jìn)行菌株復(fù)篩、耐Cd性能測定、菌種鑒定及單一介質(zhì)中基礎(chǔ)條件下其活性及Cd吸附情況研究。實(shí)驗(yàn)結(jié)果表明，菌株P(guān)DA-H具有很強(qiáng)的耐Cd能力，適宜在馬鈴薯（PDA）培養(yǎng)基中生長，不適宜在葡萄糖-蛋白胨培養(yǎng)基中生長，其在PDA固體培養(yǎng)基中能耐受的Cd濃度最高達(dá)1.4 g/L，在PDA液體培養(yǎng)基中能耐受的Cd濃度最高達(dá)0.8 g/L。生理生化鑒定及26S rDNA基因序列鑒定表明，菌株P(guān)DA-H為皺紋表膜假絲酵母（Candida rugopel -liculosa strain NRRL Y-17079）。對Cd脅迫下PDA-H在菌液中的生物量、蛋白含量、DNA含量、淀粉酶活力、脂肪酶活力及其Cd吸附能力隨時(shí)間的變化進(jìn)行考察，發(fā)現(xiàn)其分泌的淀粉酶及脂肪酶對Cd污染表現(xiàn)均很敏感，其中淀粉酶較脂肪酶更為敏感，兩種酶活性均有望作為生物指標(biāo)應(yīng)用于Cd污染的監(jiān)測中。對菌株Cd吸附能力的考察表明隨時(shí)間推移，PDA-H對Cd的吸附率總體呈不斷增加趨勢，最大吸附率達(dá)近60%。此外，Cd吸附率與脂肪酶活性呈較好的正相關(guān)關(guān)系，與淀粉酶活性呈較好的負(fù)相關(guān)關(guān)系。菌株P(guān)DA-H有望在Cd污染場地的微生物修復(fù)工程中得到應(yīng)用。 Screening and activity studies of PDA-H with high cadmium resistance Abstract At present, the heavy metal pollution of the soil is increasing. In China, soil cadmium ( Cd ) pollution is particularly remarkable, causing many hazards. Microbial remediation of heavy metal contaminated soil is a green repair technology with low cost, convenient operation and wide application prospect, many researchers all around the world pay more attention to it. This paper use a Cd resistant strain ( named PDA-H ) which was selected from soil in SuZhou typical contaminated and high organic matter site as the study object, carry out the second screening, Cd resistance determination, identification, the activity and adsorption of this strain in the single medium under the basic conditions. From these experiements, we can know PDA-H has strong resistance to Cd. This strain enjoy growing in Potato( PDA ) medium but is not suitable to live in glucose peptone mesium. The highest Cd concentration PDA-H can tolerate in solid PDA medium is 1.4 g/L, in liquid PDA medium the concentration is 0.8 g/L. From the 26S rDNA gene sequence identification, combine with physiological and biochemical identification, we can determine PDA-H is Candida rugopel -liculosa strain NRRL Y-17079. Investigating the biomass, protein content, DNA content, amylase activity, lipase activity and Cd adsorption ability of PDA-H in bacteria solution with Cd stress, it is found that amylase and lipase from PDA-H are all sensitive to Cd pollution, amylase was more sensitive than lipase. The two enzymes are expected to be used as biological indicators for the monitoring of Cd pollution. The adsorption capacity of Cd of this strain showed that the adsorption rate was increasing with the passage of time in general, and the maximum adsorption rate was close to 60%. The adsorption rate of Cd and lipase activity present positive correlation roughly, but it was just the opposite with lipase activity. Strain PDA-H is expected to be applied in microbial remediation of Cd contaminated sites. |
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