CAJ | 학술논문

目的：以不同植物中分离到的4株内生球毛壳菌NK102、NK103、NK104和NK105为对象，研究不同生态来源球毛壳菌降解木质素和纤维素的能力。方法：首先采用羧甲基纤维素和纤维素刚果红平板检测各菌株的纤维素降解能力，并利用Bavendamm平板反应检测各菌株的木质素降解能力；将4株菌分别培养在以微晶纤维素、杨树叶和木屑为惟一碳源的液体培养基中，通过检测培养液中纤维素酶和漆酶的酶活力，比较各菌株分解利用天然木质纤维素材料的能力，连续培养12 d后检测培养液中次级代谢产物的合成情况；利用已测序的球毛壳菌CBS148.51的基因组信息，寻找编码木质纤维素降解酶类的基因，为球毛壳菌分解利用木质纤维素提供分子生物学依据。结果：NK102、NK103、NK104和NK105在羧甲基纤维素培养基和纤维素刚果红培养基上都能够生长并形成水解圈；Bavendamm平板反应显示4株菌降解木质素的能力由强到弱依次是NK103、NK102、NK105和NK104。4株菌都能分解利用微晶纤维素、杨树叶和木屑，分泌纤维素酶和漆酶，其中NK102在以木屑为碳源的培养基上纤维素酶活力最强，达到0.76 U/mL发酵液，NK103在以杨树叶为碳源的培养基上漆酶活力最强。与此同时，4株菌在发酵培养过程中都能够稳定地合成球毛壳甲素（ChA)，ChA产量受到碳源影响，在以杨树叶为碳源的培养基上，NK104的ChA产量最高，可达到14.88 mg/L发酵液。利用已测序的球毛壳菌CBS148.51的基因组信息，寻找到119个编码纤维素半纤维素酶的基因、8个编码漆酶的基因和2个编码锰过氧化物酶的基因，球毛壳菌具有完整的降解纤维素半纤维素的酶体系，在木质纤维素降解真菌的开发过程中具有重要的研究价值。结论：本研究为球毛壳菌木质纤维素降解过程的研究及该菌种的开发利用奠定了基础。目的：以不同植物中分離到的4株內生毬毛殼菌NK102、NK103、NK104和NK105為對象，研究不同生態來源毬毛殼菌降解木質素和纖維素的能力。方法：首先採用羧甲基纖維素和纖維素剛果紅平闆檢測各菌株的纖維素降解能力，併利用Bavendamm平闆反應檢測各菌株的木質素降解能力；將4株菌分彆培養在以微晶纖維素、楊樹葉和木屑為惟一碳源的液體培養基中，通過檢測培養液中纖維素酶和漆酶的酶活力，比較各菌株分解利用天然木質纖維素材料的能力，連續培養12 d後檢測培養液中次級代謝產物的閤成情況；利用已測序的毬毛殼菌CBS148.51的基因組信息，尋找編碼木質纖維素降解酶類的基因，為毬毛殼菌分解利用木質纖維素提供分子生物學依據。結果：NK102、NK103、NK104和NK105在羧甲基纖維素培養基和纖維素剛果紅培養基上都能夠生長併形成水解圈；Bavendamm平闆反應顯示4株菌降解木質素的能力由彊到弱依次是NK103、NK102、NK105和NK104。4株菌都能分解利用微晶纖維素、楊樹葉和木屑，分泌纖維素酶和漆酶，其中NK102在以木屑為碳源的培養基上纖維素酶活力最彊，達到0.76 U/mL髮酵液，NK103在以楊樹葉為碳源的培養基上漆酶活力最彊。與此同時，4株菌在髮酵培養過程中都能夠穩定地閤成毬毛殼甲素（ChA)，ChA產量受到碳源影響，在以楊樹葉為碳源的培養基上，NK104的ChA產量最高，可達到14.88 mg/L髮酵液。利用已測序的毬毛殼菌CBS148.51的基因組信息，尋找到119箇編碼纖維素半纖維素酶的基因、8箇編碼漆酶的基因和2箇編碼錳過氧化物酶的基因，毬毛殼菌具有完整的降解纖維素半纖維素的酶體繫，在木質纖維素降解真菌的開髮過程中具有重要的研究價值。結論：本研究為毬毛殼菌木質纖維素降解過程的研究及該菌種的開髮利用奠定瞭基礎。
목적：이불동식물중분리도적4주내생구모각균NK102、NK103、NK104화NK105위대상，연구불동생태래원구모각균강해목질소화섬유소적능력。방법：수선채용최갑기섬유소화섬유소강과홍평판검측각균주적섬유소강해능력，병이용Bavendamm평판반응검측각균주적목질소강해능력；장4주균분별배양재이미정섬유소、양수협화목설위유일탄원적액체배양기중，통과검측배양액중섬유소매화칠매적매활력，비교각균주분해이용천연목질섬유소재료적능력，련속배양12 d후검측배양액중차급대사산물적합성정황；이용이측서적구모각균CBS148.51적기인조신식，심조편마목질섬유소강해매류적기인，위구모각균분해이용목질섬유소제공분자생물학의거。결과：NK102、NK103、NK104화NK105재최갑기섬유소배양기화섬유소강과홍배양기상도능구생장병형성수해권；Bavendamm평판반응현시4주균강해목질소적능력유강도약의차시NK103、NK102、NK105화NK104。4주균도능분해이용미정섬유소、양수협화목설，분비섬유소매화칠매，기중NK102재이목설위탄원적배양기상섬유소매활력최강，체도0.76 U/mL발효액，NK103재이양수협위탄원적배양기상칠매활력최강。여차동시，4주균재발효배양과정중도능구은정지합성구모각갑소（ChA)，ChA산량수도탄원영향，재이양수협위탄원적배양기상，NK104적ChA산량최고，가체도14.88 mg/L발효액。이용이측서적구모각균CBS148.51적기인조신식，심조도119개편마섬유소반섬유소매적기인、8개편마칠매적기인화2개편마맹과양화물매적기인，구모각균구유완정적강해섬유소반섬유소적매체계，재목질섬유소강해진균적개발과정중구유중요적연구개치。결론：본연구위구모각균목질섬유소강해과정적연구급해균충적개발이용전정료기출。
Objective: The decomposition abilities of lignocellulosic materials by Chaetomium globosum NK102, NK103, NK104 and NK105, endophytes from different plants were evaluated. Methods: The cellulose utilizing ca-pability of the C.globosum isolates were tested on carboxymethylcellulose agar and cellulose-congo red agar. The lignin utilizing experiments were performed on Bavendamm plates, and the degradative activity was compared by measuring the respective zone of color change. The lignocellulases production potential of these isolates using mi-crocrystalline cellulose, the leaves of Populus sp. and wood powder as the sole carbon source in liquid fermenta-tion was assessed. In addition, secondary metabolites produced by the C.globosum isolates were detected after 12 days cultivation. In the sequenced genome of C.globosum CBS148.51, genes encoding enzymes involved in lignocel-luloses degrading were identified by sequence homology alignment. Results: C.globosum NK102, NK103, NK104 and NK105 formed clear zones when cultured on carboxymethylcellulose or cellulose-congo red agar. In addition, all four isolates exhibited biodegradation capabilities against lignin and the strong-to-weak sequence was NK 103, NK102, NK105 and NK104 by Bavendamm reaction. In liquid culture, all isolates secreted cellulases and laccase on agar containing microcrystalline cellulose, the leaves of Populus sp., and wood powder. The highest activity of cellulase(0.76 U/mL) was obtained by NK102 when cultured on wood powder medium supplemented with peptone. The highest activity of laccase was obtained by NK103 when cultured on the leaves of Populus sp. medium. Chae-toglobosin A(ChA) was detected in the cultue broth of all four isolates. The yield of ChA was affected by carbon biomass and the highest yield was obtained by NK104 when cultured on the leaves of Populus with a yield of 14.88 mg/L. In the sequenced genome of C.globosum CBS148.51, we defined 119 genes encoding enzymes in-volved in cellulose and hemicellulose degradation, 8 genes for laccase and 2 genes for manganese peroxidase by sequence homology alignment, suggesting that C.globosum had a complete enzyme system on lignocelluloses utiliza-tion and deserved for further study for possible industrial and environmental application. Conclusion: This study shed lights on the molecular mechanism of lignocellulose-degrading in C.globosum and provided information for strain screening.