Kamande Stephen M, Omwenga George I, Ngugi Mathew P
Technical University of Kenya, Department of Biochemistry and Biotechnology, P.O. Box 52428-00200, Nairobi, Kenya.
Kenyatta University, Department of Biochemistry, Biotechnology and Microbiology, P.O. Box 43844-00100, Nairobi, Kenya.
Heliyon. 2024 Aug 30;10(17):e36802. doi: 10.1016/j.heliyon.2024.e36802. eCollection 2024 Sep 15.
Two species of (KM01, KM03) and sp.KM02 isolated from decaying plant biomass were evaluated for their ability to produce cellulases on maize cob, eucalyptus, and cypress substrates under solid-state fermentation. A total of 10 fungal samples from decaying plant biomass were collected from Karura forest based on morphological variations. The fungi isolated were screened for cellulase activity and positive isolates were selected for the study. ITS4 R and ITS86 F primers were used to identify the fungal isolates with accuracy ranging from 98 % to 100 %. The crude cellulases produced was assayed for FPase, exoglucanase, endoglucanase and β-glucosidase. Cellulases of sp. KM01 produced higher FPase and exoglucanase (2.01 ± 0.13 IU/ml and 0.94 ± 0.08 IU/ml) on pretreated maize cobs with 0.1M HCl at 121C, while that of sp.KM03 produced higher β-glucosidase and endoglucanase (588.6 ± 64.2 IU/ml and 3.59 ± 0.02 IU/ml) on maize cobs pretreated with 0.1M NaOH at 121C. However, cellulases of sp. KM01 produced higher β-glucosidase and FPase (629.7 ± 20.2 IU/ml and 1.67 ± 0.03 IU/ml) on untreated maize cobs after the 9th day of incubation, whereas cellulases of sp.KM03 and sp.KM02 produced higher endoglucanase and exoglucanase (2.80 ± 0.21 IU/ml and 0.83 ± 0.02 IU/ml) on untreated maize cobs after the 3rd and 6th day of incubation. Saccharification of maize cobs by cellulase of sp.KM03 produced the highest reducing sugars at 8 % substrate loading (10.17 ± 0.37 mg/ml) after 72 h of incubation. Simultaneous hydrolysis and fermentation of maize cobs by cellulase of sp.KM02 and yielded higher bioethanol (28.72 ± 3.82 mg/ml) after 96 h of fermentation. Maize cob is established as a suitable feedstock for cellulases and bioethanol production.
对从腐烂植物生物质中分离出的两种曲霉属真菌(KM01、KM03)和一种未鉴定的曲霉属真菌KM02,在固态发酵条件下,评估它们在玉米芯、桉木和柏木底物上产生纤维素酶的能力。基于形态学差异,从卡鲁拉森林中收集了总共10个来自腐烂植物生物质的真菌样本。对分离出的真菌进行纤维素酶活性筛选,并选择阳性分离株进行研究。使用ITS4 R和ITS86 F引物准确鉴定真菌分离株,准确率在98%至100%之间。对所产生的粗纤维素酶进行滤纸酶活性(FPase)、外切葡聚糖酶、内切葡聚糖酶和β - 葡萄糖苷酶的测定。曲霉属真菌KM01的纤维素酶在121℃下用0.1M盐酸预处理的玉米芯上产生较高的FPase和外切葡聚糖酶(2.01±0.13 IU/ml和0.94±0.08 IU/ml),而曲霉属真菌KM03的纤维素酶在121℃下用0.1M氢氧化钠预处理的玉米芯上产生较高的β - 葡萄糖苷酶和内切葡聚糖酶(588.6±64.2 IU/ml和3.59±0.02 IU/ml)。然而,曲霉属真菌KM01的纤维素酶在培养第9天后,在未处理的玉米芯上产生较高的β - 葡萄糖苷酶和FPase(629.7±20.2 IU/ml和1.67±0.03 IU/ml),而曲霉属真菌KM03和KM02的纤维素酶在培养第3天和第6天后,在未处理的玉米芯上产生较高的内切葡聚糖酶和外切葡聚糖酶(2.80±0.21 IU/ml和0.83±0.02 IU/ml)。曲霉属真菌KM03的纤维素酶在72小时培养后,在底物负载量为8%时糖化玉米芯产生的还原糖最高(10.17±0.37 mg/ml)。曲霉属真菌KM0