水解酶活性在秸秆静态高温堆腐过程中变化的研究
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摘要
农业废弃物的堆腐是在微生物作用下,通过发酵使有机物矿质化、腐殖化和无害化而变成腐熟肥料的过程。在微生物分解有机物的过程中,不但可以生成大量可被植物吸收利用的有效态氮、磷、钾化合物,还会合成新的高分子有机物—腐殖质。由于这一过程是在微生物分泌酶的作用下进行的酶促反应过程,研究酶活性变化可以从本质上揭示堆腐过程的生物作用与机理,对秸秆的有效利用有着重要的科学价值。本研究以小麦秸秆、玉米秸秆和尿素为堆腐材料,利用静态高温堆腐装置,系统性研究了堆腐过程纤维素酶、蔗糖酶、脲酶、蛋白酶活性以及堆腐过程温度、含水率、电导率(EC)、pH值变化规律。
以小麦秸秆为原料进行堆肥试验,分别进行添加葡萄糖活化菌剂(G+J)、添加菌剂(J)、不添加菌剂(CK)三种处理;以玉米秸秆为原料进行堆肥试验,进行添加微生物腐解菌剂(J)和不添加菌剂(CK)两种处理。实验结果表明:
1.在以小麦秸秆和尿素为堆料的腐解试验中,G+J处理的纤维素酶活性峰值出现在第9d,达到334.37mg glucose(g.24h)~(-1),J和CK酶活性峰值较G+J滞后3d和6d出现,酶活性峰值分别为271.59mg glucose(g.24h)~(-1)和236.67mg glucose(g.24h)~(-1),添加微生物菌剂使得堆体纤维素酶活性变化趋势发生很大变异;在以玉米秸秆和尿素为堆料的腐解试验中,J处理的纤维素酶活性峰值出现在第9d为352.11mg glucose(g.24h)~(-1),CK处理的酶活性峰值较J处理滞后3d,为285.51mg glucose(g.24h)~(-1),加入微生物菌剂后纤维素酶在堆腐过程中的变化趋势同CK处理基本一致。
2.在以小麦秸秆和尿素为堆料的腐解试验中,G+J处理的蔗糖酶活性在第9d达到最高峰值,为1582.64 mg glucose(g.24h)~(-1),J在第12d达到峰值,为1455.18mgglucose(g.24h)~(-1),CK处理的蔗糖酶活性峰值出现在第15d,为625.06 mg glucose(g.24h)~(-1),G+J处理蔗糖酶活性峰值是CK处理的2.53倍,J处理蔗糖酶活性是CK处理的2.33倍,表明加入菌剂能够提高蔗糖酶活性,微生物菌剂可以加速农业废弃物的矿化;在以玉米秸秆和尿素为堆料的腐解试验中,堆腐中期4d到12d两个处理的温度和蔗糖酶活性变化趋势一致,随着温度的降低,蔗糖酶活性降低。
3.在以小麦秸秆和尿素为堆料的腐解试验中,在堆腐23d后各处理脲酶活性均小于20.00mg NH_3(g.24h)~(-1)。3个处理整个堆腐过程脲酶活性的平均值分别为50.69±26.17 mgNH_3(g.24h)~(-1)、34.68±7.58 mg NH_3(g.24h)~(-1)和25.20±5.08 mg NH_3(g.24h)~(-1),添加复合微生物菌剂可使G+J和J处理在整个堆腐过程中脲酶活性均高于不接菌的CK;在以玉米秸秆和尿素为堆料的腐解试验中,添加复合微生物菌剂后,整个堆腐过程中脲酶平均活性为(215.50±49.96)mg NH_3(g.24h)~(-1),显著高于不接菌的CK(F=18.73,P＜0.05)。可见接种复合微生物菌剂可以提高堆料堆腐过程中脲酶活性,加速酰胺化合物的转化。
4.在以小麦秸秆和尿素为堆料的腐解试验中,温度在第2d迅速升高,而酶活性降低,可见高温对蛋白酶活性有抑制作用。在以玉米秸秆和尿素为堆料的腐解试验中,添加复合微生物菌剂后,整个堆腐过程中平均蛋白酶活性为(9.63±1.37)mg NH_2(g.24h)~(-1),而不接菌的CK则为(8.38±0.98)mg NH_2(g.24h)~(-1),变异系数分别为14.23%和11.69%,但是两个处理间差异不显著(P＞0.05),原因可能是由于玉米秸秆本身粗蛋白质含量很少,因而诱导其分泌的底物含量少。
5.小麦秸秆和尿素堆腐过程,G+J、J和CK处理的最高温度分别为66℃、67.1℃和59.5℃,整个堆腐过程温度大于50℃的时间为:G+J(240 h)＞J(120 h)＞CK(96 h);玉米秸秆和尿素堆腐过程,J和CK处理的最高温度分别为73.3℃、65.9℃,整个堆腐过程温度大于50℃的时间为:J(240 h)＞CK(192 h);小麦秸秆、玉米秸秆和尿素整个堆腐过程含水率都呈下降趋势,;小麦秸秆和尿素堆腐起始阶段EC值上升,在堆腐12d 3个处理达到高峰值分别为2.69 ms/cm、2.32 ms/cm、2.30 ms/cm,随后EC值下降,玉米秸秆和尿素从堆腐第2d开始到结束,接菌处理的EC值均高于不接菌的对照CK,整个堆腐过程总的EC值分别为26.59ms/cm和23.26ms/cm;小麦秸秆和尿素堆腐过程pH值在第2d升高,在第4d下降,随后呈上升趋势,玉米秸秆和尿素堆腐过程变化趋势同小麦秸秆。两种不同物料加入微生物菌剂处理的pH值变化幅度较小,且pH值相对较低。
The compost of agricultural waste materials is the process that under the function of microorganism,the organic matter turn into well-rotted manure by the way of the organic matter being mineralized and humus and harmless.In the process of the function of microorganism,not only massive the nitrogen,phosphorus and potassium compounds that are useful to plant are produced,also new high polymer organic matter---humus is synthesized.
Because the process is the reaction of enzyme pressing,this is produced by microorganism,
study the changes of enzymes activity is helpful to explain the process and mechanism of composting well and offer important scientific value.The changes of the activity of the cellulose,sucrose,urea enzymes,protein enzyme,temperature,water content,electric conductivity,and pH had been studied during an aeration composting process in a static state using substrate of mixed wheat straw,maize straw and urea with and without adding microorganism agent.
Three treatments of adding microorganism agents actived glucose(G+J),adding microorganism agents(J) and without microorganism agents(CK) were made in wheat straw and urea composting process.Two treatments of adding microorganism agents(J) and without microorganism agents(CK) were made in maize straw and urea composting process. The main results are showed as follows:
1.The peak value of the cellulose enzyme activity in the treatment of G+J was observed in 9 d,and was earlier than J and CK for 3 d and 6 d,among them the values were 334.37 mg glucose(g.24h)~(-1),271.59 mg glucose(g.24h)~(-1) and 236.67 mg glucose(g.24h)~(-1) respectively during the wheat straw and urea composting process.There was a large variation in trends of cellulose enzyme activity.While during the maize straw and urea composting process,the peak value of the cellulose enzyme activity in the treatment of J was observed in 9 d,and was earlier than CK for 3 d,among them the values were 352.11 mg glucose(g.24h)~(-1) and 285.51 mg glucose(g.24h)~(-1) respectively.The trend of cellulose enzyme activity adding microorganism agents was the same as that without microorganism agents.
2.The peak value of the sucrose enzyme activity in the treatment of G+J was observed in 9 d,and was earlier than J and CK for 3 d and 6 d,among them the values were 1582.64 mg glucose(g.24h)~(-1),1455.18 mg glucose(g.24h)~(-1) and 625.06 mg glucose(g.24h)~(-1) respectively during the wheat straw and urea composting process.And the peak values of sucrose enzyme activity in the treatments of G+J and J were 2.53 times and 2.33 times than CK,respectively.It represents that adding microorganism agents can enhance the sucrose enzyme activity and accelerate the mineralization of agricultural wastes.While during the maize straw and urea composting process,the trend of sucrose enzyme activity was in accordance with the temperature during the period of 4 d and 12 d,and the sucrose enzyme activity fell with the decrease of temperature.
3.The urea enzyme activities of three treatments were lower than 20.00 mg NH_3 (g.24h)~(-1) after 23 d during the wheat straw and urea composting process.The average activities of urea enzyme of three treatments were 50.69±26.17 mg NH_3(g.24h)~(-1),34.68±7.58 mg NH_3(g.24h)~(-1) and 25.20±5.08 mg NH_3(g.24h)~(-1),respectively.Adding microorganism agents can enhance the urea enzyme activities.The average urea enzyme activity was (215.50±49.96) mg NH_3(g.24h)~(-1) in treatment of J in maize straw and urea composting process,much higher than CK(F=18.73,P＜0.05).It illustrates that adding microorganism agents can enhance the urea enzyme activity and accelerate transformation of amide compounds.
4.In the wheat straw and urea composting process,the activities of protein enzyme decreased with rise of temperature in 2 d in three treatments,so,high temperature can restrain protein enzyme activities.The average activities of protein enzyme were(9.63±1.37) NH_2(g.24h)~(-1) and(8.38±0.98) NH_2(g.24h)~(-1) in treatment of J and CK during maize straw and urea composting process.And the coefficient of variation were 14.23%11.69%,respectively. The reason maybe that maize straw had little raw protein,so it can induce little substrate.
5.The highest temperature for G+J、J and CK were 66℃、67.1℃and 59.5℃, respectively.The rank of time for temperature higher than 50℃was:G+J(240 h)＞J(120 h)＞CK(96 h) during the wheat straw and urea composting process.The highest temperature were 73.3℃and 65.9℃for J and CK and the rank of time for temperature higher than 50℃was:＞J(240 h)＞CK(192 h) during the maize straw and urea composting process.Water content decreased in wheat and maize straw and urea composting process.The values of EC were 2.69 ms/cm,2.32 ms/cm and 2.30 ms/cm for three treatments in 12 d during the wheat straw and urea composting process.The value of EC of the treatment adding microbial inoculants was higher than that without microbial inoculants during the maize straw and urea composting process.And the values of EC were 26.59 ms/cm and 23.26 ms/cm.The value of pH increased in 2d,and decreased in 4d,and then on the rise in wheat straw and urea composting process.The same situation was detected in maize and urea composting process. Lower pH values and small range of changes in pH values were founded in two different agricultural materials adding microbial inoculants.
以小麦秸秆为原料进行堆肥试验,分别进行添加葡萄糖活化菌剂(G+J)、添加菌剂(J)、不添加菌剂(CK)三种处理;以玉米秸秆为原料进行堆肥试验,进行添加微生物腐解菌剂(J)和不添加菌剂(CK)两种处理。实验结果表明:
1.在以小麦秸秆和尿素为堆料的腐解试验中,G+J处理的纤维素酶活性峰值出现在第9d,达到334.37mg glucose(g.24h)~(-1),J和CK酶活性峰值较G+J滞后3d和6d出现,酶活性峰值分别为271.59mg glucose(g.24h)~(-1)和236.67mg glucose(g.24h)~(-1),添加微生物菌剂使得堆体纤维素酶活性变化趋势发生很大变异;在以玉米秸秆和尿素为堆料的腐解试验中,J处理的纤维素酶活性峰值出现在第9d为352.11mg glucose(g.24h)~(-1),CK处理的酶活性峰值较J处理滞后3d,为285.51mg glucose(g.24h)~(-1),加入微生物菌剂后纤维素酶在堆腐过程中的变化趋势同CK处理基本一致。
2.在以小麦秸秆和尿素为堆料的腐解试验中,G+J处理的蔗糖酶活性在第9d达到最高峰值,为1582.64 mg glucose(g.24h)~(-1),J在第12d达到峰值,为1455.18mgglucose(g.24h)~(-1),CK处理的蔗糖酶活性峰值出现在第15d,为625.06 mg glucose(g.24h)~(-1),G+J处理蔗糖酶活性峰值是CK处理的2.53倍,J处理蔗糖酶活性是CK处理的2.33倍,表明加入菌剂能够提高蔗糖酶活性,微生物菌剂可以加速农业废弃物的矿化;在以玉米秸秆和尿素为堆料的腐解试验中,堆腐中期4d到12d两个处理的温度和蔗糖酶活性变化趋势一致,随着温度的降低,蔗糖酶活性降低。
3.在以小麦秸秆和尿素为堆料的腐解试验中,在堆腐23d后各处理脲酶活性均小于20.00mg NH_3(g.24h)~(-1)。3个处理整个堆腐过程脲酶活性的平均值分别为50.69±26.17 mgNH_3(g.24h)~(-1)、34.68±7.58 mg NH_3(g.24h)~(-1)和25.20±5.08 mg NH_3(g.24h)~(-1),添加复合微生物菌剂可使G+J和J处理在整个堆腐过程中脲酶活性均高于不接菌的CK;在以玉米秸秆和尿素为堆料的腐解试验中,添加复合微生物菌剂后,整个堆腐过程中脲酶平均活性为(215.50±49.96)mg NH_3(g.24h)~(-1),显著高于不接菌的CK(F=18.73,P＜0.05)。可见接种复合微生物菌剂可以提高堆料堆腐过程中脲酶活性,加速酰胺化合物的转化。
4.在以小麦秸秆和尿素为堆料的腐解试验中,温度在第2d迅速升高,而酶活性降低,可见高温对蛋白酶活性有抑制作用。在以玉米秸秆和尿素为堆料的腐解试验中,添加复合微生物菌剂后,整个堆腐过程中平均蛋白酶活性为(9.63±1.37)mg NH_2(g.24h)~(-1),而不接菌的CK则为(8.38±0.98)mg NH_2(g.24h)~(-1),变异系数分别为14.23%和11.69%,但是两个处理间差异不显著(P＞0.05),原因可能是由于玉米秸秆本身粗蛋白质含量很少,因而诱导其分泌的底物含量少。
5.小麦秸秆和尿素堆腐过程,G+J、J和CK处理的最高温度分别为66℃、67.1℃和59.5℃,整个堆腐过程温度大于50℃的时间为:G+J(240 h)＞J(120 h)＞CK(96 h);玉米秸秆和尿素堆腐过程,J和CK处理的最高温度分别为73.3℃、65.9℃,整个堆腐过程温度大于50℃的时间为:J(240 h)＞CK(192 h);小麦秸秆、玉米秸秆和尿素整个堆腐过程含水率都呈下降趋势,;小麦秸秆和尿素堆腐起始阶段EC值上升,在堆腐12d 3个处理达到高峰值分别为2.69 ms/cm、2.32 ms/cm、2.30 ms/cm,随后EC值下降,玉米秸秆和尿素从堆腐第2d开始到结束,接菌处理的EC值均高于不接菌的对照CK,整个堆腐过程总的EC值分别为26.59ms/cm和23.26ms/cm;小麦秸秆和尿素堆腐过程pH值在第2d升高,在第4d下降,随后呈上升趋势,玉米秸秆和尿素堆腐过程变化趋势同小麦秸秆。两种不同物料加入微生物菌剂处理的pH值变化幅度较小,且pH值相对较低。
The compost of agricultural waste materials is the process that under the function of microorganism,the organic matter turn into well-rotted manure by the way of the organic matter being mineralized and humus and harmless.In the process of the function of microorganism,not only massive the nitrogen,phosphorus and potassium compounds that are useful to plant are produced,also new high polymer organic matter---humus is synthesized.
Because the process is the reaction of enzyme pressing,this is produced by microorganism,
study the changes of enzymes activity is helpful to explain the process and mechanism of composting well and offer important scientific value.The changes of the activity of the cellulose,sucrose,urea enzymes,protein enzyme,temperature,water content,electric conductivity,and pH had been studied during an aeration composting process in a static state using substrate of mixed wheat straw,maize straw and urea with and without adding microorganism agent.
Three treatments of adding microorganism agents actived glucose(G+J),adding microorganism agents(J) and without microorganism agents(CK) were made in wheat straw and urea composting process.Two treatments of adding microorganism agents(J) and without microorganism agents(CK) were made in maize straw and urea composting process. The main results are showed as follows:
1.The peak value of the cellulose enzyme activity in the treatment of G+J was observed in 9 d,and was earlier than J and CK for 3 d and 6 d,among them the values were 334.37 mg glucose(g.24h)~(-1),271.59 mg glucose(g.24h)~(-1) and 236.67 mg glucose(g.24h)~(-1) respectively during the wheat straw and urea composting process.There was a large variation in trends of cellulose enzyme activity.While during the maize straw and urea composting process,the peak value of the cellulose enzyme activity in the treatment of J was observed in 9 d,and was earlier than CK for 3 d,among them the values were 352.11 mg glucose(g.24h)~(-1) and 285.51 mg glucose(g.24h)~(-1) respectively.The trend of cellulose enzyme activity adding microorganism agents was the same as that without microorganism agents.
2.The peak value of the sucrose enzyme activity in the treatment of G+J was observed in 9 d,and was earlier than J and CK for 3 d and 6 d,among them the values were 1582.64 mg glucose(g.24h)~(-1),1455.18 mg glucose(g.24h)~(-1) and 625.06 mg glucose(g.24h)~(-1) respectively during the wheat straw and urea composting process.And the peak values of sucrose enzyme activity in the treatments of G+J and J were 2.53 times and 2.33 times than CK,respectively.It represents that adding microorganism agents can enhance the sucrose enzyme activity and accelerate the mineralization of agricultural wastes.While during the maize straw and urea composting process,the trend of sucrose enzyme activity was in accordance with the temperature during the period of 4 d and 12 d,and the sucrose enzyme activity fell with the decrease of temperature.
3.The urea enzyme activities of three treatments were lower than 20.00 mg NH_3 (g.24h)~(-1) after 23 d during the wheat straw and urea composting process.The average activities of urea enzyme of three treatments were 50.69±26.17 mg NH_3(g.24h)~(-1),34.68±7.58 mg NH_3(g.24h)~(-1) and 25.20±5.08 mg NH_3(g.24h)~(-1),respectively.Adding microorganism agents can enhance the urea enzyme activities.The average urea enzyme activity was (215.50±49.96) mg NH_3(g.24h)~(-1) in treatment of J in maize straw and urea composting process,much higher than CK(F=18.73,P＜0.05).It illustrates that adding microorganism agents can enhance the urea enzyme activity and accelerate transformation of amide compounds.
4.In the wheat straw and urea composting process,the activities of protein enzyme decreased with rise of temperature in 2 d in three treatments,so,high temperature can restrain protein enzyme activities.The average activities of protein enzyme were(9.63±1.37) NH_2(g.24h)~(-1) and(8.38±0.98) NH_2(g.24h)~(-1) in treatment of J and CK during maize straw and urea composting process.And the coefficient of variation were 14.23%11.69%,respectively. The reason maybe that maize straw had little raw protein,so it can induce little substrate.
5.The highest temperature for G+J、J and CK were 66℃、67.1℃and 59.5℃, respectively.The rank of time for temperature higher than 50℃was:G+J(240 h)＞J(120 h)＞CK(96 h) during the wheat straw and urea composting process.The highest temperature were 73.3℃and 65.9℃for J and CK and the rank of time for temperature higher than 50℃was:＞J(240 h)＞CK(192 h) during the maize straw and urea composting process.Water content decreased in wheat and maize straw and urea composting process.The values of EC were 2.69 ms/cm,2.32 ms/cm and 2.30 ms/cm for three treatments in 12 d during the wheat straw and urea composting process.The value of EC of the treatment adding microbial inoculants was higher than that without microbial inoculants during the maize straw and urea composting process.And the values of EC were 26.59 ms/cm and 23.26 ms/cm.The value of pH increased in 2d,and decreased in 4d,and then on the rise in wheat straw and urea composting process.The same situation was detected in maize and urea composting process. Lower pH values and small range of changes in pH values were founded in two different agricultural materials adding microbial inoculants.
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