Sharma K K, Kapoor M, Kuhad R C
Department of Microbiology, University of Delhi South Campus, New Delhi, India.
Lett Appl Microbiol. 2005;41(1):24-31. doi: 10.1111/j.1472-765X.2005.01721.x.
The effect of in vivo enzymatic digestion (IVED), in vitro xylanase digestion (IVXD), metabolic analogues, surfactants and polyethylene glycol (PEG) on laccase production from Ganoderma sp. kk-02 was studied.
An acidic laccase producing Ganoderma sp. kk-02 produced 16.0 U ml(-1) and 365.0 U g(-1) of laccase, when grown under submerged (SmF) and solid state (SSF) fermentation conditions, respectively. Modification of the substrate (wheat bran) molecular architecture by IVED and IVXD increased subsequent laccase production from Ganoderma sp. kk-02 by 1.31-fold (21.0 U ml(-1)) (SmF); 2.21-fold (810.0 U g(-1)) (SSF) and 1.10-fold (18.0 U ml(-1)) (SmF); 1.78-fold (650.0 U g(-1)) (SSF) when compared with untreated wheat bran. Further enhancement in laccase yield under SmF and SSF was obtained when IVED treated wheat bran was used in conjunction with amino acids [DL-tryptophan, 2.66-fold (56.0 U ml(-1)) SmF; 2.86-fold (2324.0 U g(-1)) SSF], vitamins [biotin, 1.71-fold (36.0 U ml(-1)) SmF; 3.06-fold (2483.0 U g(-1)) SSF], surfactants [Tween-40, 1.85-fold (39.0 U ml(-1)) SmF; 2.25-fold (1828.0 U g(-1)) SSF], and PEG [PEG 6000, 1.93-fold (40.0 U ml(-1)) SmF; 1.58-fold (1284.0 U g(-1)) SSF].
The IVED of substrate (wheat bran) facilitated hyper laccase production in presence of additives from Ganoderma sp. kk-02.
The study highlights a new methodology viz. IVED for concomitant and economic production of diverse enzymes using the same substrate. The hyper laccase levels obtained could improve the economic competitiveness of environmentally benign processes applied in varied industries. The work also provides an insight into the regulation of complex metabolic pathways governing the expression of extra cellular proteins from white-rot fungi.
研究体内酶消化（IVED）、体外木聚糖酶消化（IVXD）、代谢类似物、表面活性剂和聚乙二醇（PEG）对灵芝kk - 02产漆酶的影响。
一株产酸性漆酶的灵芝kk - 02在液体深层发酵（SmF）和固态发酵（SSF）条件下生长时，分别产生16.0 U ml⁻¹和365.0 U g⁻¹的漆酶。通过IVED和IVXD对底物（麦麸）分子结构进行修饰，与未处理的麦麸相比，灵芝kk - 02随后的漆酶产量在SmF条件下提高了1.31倍（21.0 U ml⁻¹），在SSF条件下提高了2.21倍（810.0 U g⁻¹）；在SmF条件下提高了1.10倍（18.0 U ml⁻¹），在SSF条件下提高了1.78倍（650.0 U g⁻¹）。当将IVED处理的麦麸与氨基酸[DL - 色氨酸，在SmF条件下提高2.66倍（56.0 U ml⁻¹）；在SSF条件下提高2.86倍（2324.0 U g⁻¹）]、维生素[生物素，在SmF条件下提高1.71倍（36.0 U ml⁻¹）；在SSF条件下提高3.06倍（2483.0 U g⁻¹）]、表面活性剂[吐温 - 40，在SmF条件下提高1.85倍（39.0 U ml⁻¹）；在SSF条件下提高2.25倍（1828.0 U g⁻¹）]和PEG[PEG 6000，在SmF条件下提高1.93倍（40.0 U ml⁻¹）；在SSF条件下提高1.58倍（1284.0 U g⁻¹）]联合使用时，SmF和SSF条件下的漆酶产量进一步提高。
底物（麦麸）的IVED在添加剂存在的情况下促进了灵芝kk - 02高产漆酶。
该研究突出了一种新方法，即IVED可利用同一底物同时且经济地生产多种酶。所获得的高产漆酶水平可提高应用于不同行业的环境友好型工艺的经济竞争力。该工作还深入了解了控制白腐真菌细胞外蛋白质表达的复杂代谢途径的调控机制。
