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第 5 期 姚炎 等:溜曲霉 M3b 产壳聚糖酶的固体发酵工艺优化及酶学性质 221
接种量太低,菌的浓度低,生长缓慢,酶活力低;接种 j.carbpol.2021.117799.
量太高,造成固体发酵基质含水量高,自由水含量 [3] GUO N,SUN J N,WANG W,et al. Cloning,expression and
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体交换,影响散热,不利于菌株的生长。一般固体发 [4] 张馨月,岳晓洁,李铮峥,等 .淡紫紫孢菌的筛选、鉴定及产壳聚
酵初始含水量在30%~70%。 糖酶固体发酵条件优化[J]. 中国食品学报,2020,20(4):160-
169.ZHANG X Y,YUE X J,LI Z Z,et al.Isolation and identifi⁃
本研究在单因素试验基础上,对固体发酵的温
cation of Purpureocillium lilacinum and optimization of its solid
度、时间、起始 pH 进行了优化,结果显示,最优组合 fermentation conditions for producing chitosanase[J]. Journal of
为发酵 温 度 32 ℃ 、起 始 pH 7.0、发 酵 时 间 6 d,在 Chinese institute of food science and technology,2020,20(4):
此条件下 M3b 产壳聚糖酶水平达到 20.56 U/mL。 160-169(in Chinese with English abstract).
[5] 李佳茵,王慧敏,祖国仁 .海洋真菌 MF-08产壳聚糖酶诱导条件
这 与 采 用 固 体 发 酵 产 壳 聚 糖 酶 鹿 皮 曲 霉(8.26
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olyporum(0.61 U/mL) 相比,表明溜曲霉 M3b 固
[15]
1781(in Chinese with English abstract).
体发酵产壳聚糖酶活力更高且具有优势。 [6] 曹永强,余志坚,陈超,等 . 一株嗜热链球菌乳糖酶的分离纯化
不同菌株产壳聚糖酶的最适温度不同。本研究 及酶学特性研究[J]. 食品科学技术学报,2018,36(2):42-51.
中菌株M3b产壳聚糖酶的最适温度为60 ℃。一般来 CAO Y Q,YU Z J,CHEN C,et al.Isolation,purification and en⁃
zyme characteristics of lactase produced by a Streptococcus ther⁃
说,真菌壳聚糖酶的最适温度在 50~55 ℃,Fusarium
mophiles strain[J].Journal of food science and technology,2018,
solani 最适温度为 40 ℃,Penicilliumi slandicum 最适 36(2):42-51(in Chinese with English abstract).
温度为 45 ℃,Aspergillus oryzae IAM2660 最适温度 [7] FU X,GUO Y X,JIN Y G,et al.Bioconversion of chitin waste
为 50 ℃ [16] 。自然界中大部分壳聚糖酶不耐热。因 using a cold-adapted chitinase to produce chitin oligosaccharides
[J/OL]. LWT,2020,133:109863[2022-04-07]. https://doi.
此,溜曲霉壳聚糖酶相对于其他真菌壳聚糖酶更适 org/10.1016/j.lwt.2020.109863.
合工业生产。菌株 M3b 产壳聚糖酶的最适 pH 为 [8] JIANG Z Q,MAּ S A,GUAN LY,et al.Biochemical character⁃
5.5,这与之前报道的托球菌属 JG [17] 和曲霉 QD-2 [18] ization of a novel bifunctional chitosanase from Paenibacillus
barengoltzii for chitooligosaccharide production[J/OL]. World
产壳聚糖酶在 pH 5.6 时酶活力最高相一致。海洋真
journal of microbiology and biotechnology,2021,37(5):1-13
菌 MF-08 最适 pH 5.2,但 pH 在 4.0~6.0 才相对稳 [2022-04-07].https://doi.org/10.1007/s11274-021-03051-0 83.
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定 。Aspergillus sp. CJ22-326产生的壳聚糖酶的最 [9] DOAN C T,TRAN T N,NGUYEN V B,et al.Production of a
thermostable chitosanase from shrimp heads via Paenibacillus
适 pH 4.0,pH 在 3.5~7.5 的范围内保持稳定 [19] 。一
mucilaginosus TKU032 conversion and its application in the
般情况下,真菌较细菌的稳定范围小,而溜曲霉产真 preparation of bioactive chitosan oligosaccharides[J/OL].Marine
菌壳聚糖酶在较广的 pH 范围(3.0~9.0)保持相对稳 drugs,2019,17(4):217[2022-04-07].https://doi.org/10.3390/
定,宽泛的酸碱适应范围使其具备优异的工业化发 md17040217.
[10] DING M,ZHANG T,SUN C,et al.A Chitosanase mutant from
酵优势。壳聚糖酶 AtChito40 水解壳聚糖最后得到
Streptomyces sp. N174 prefers to produce functional chitopenta⁃
产物的聚合度≥2,最小酶解产物为壳二糖。这与类 saccharide[J].International journal of biological macromolecules,
[20] 2020,151:1091-1098.
芽孢杆菌 TKU047 产壳聚糖酶的水解产物在反应
初期仅产生聚合度大于 2 的壳寡糖但没有氨基葡萄 [11] EMBABY A M,MELIKA R R,HUSSEIN A,et al.Biosynthe⁃
sis of chitosan-oligosaccharides(COS)by non-aflatoxigenic As⁃
糖(GlcN)的结果一致,表明壳聚糖酶 AtChito40是一 pergillus sp.strain EGY1 DSM 101520:a robust biotechnologi⁃
种内切酶,能更加有效地降解壳聚糖。 cal approach[J].Process biochemistry,2018,64:16-30.
[12] 冯志彬,薛钰,陈国忠,等 .1 株产壳聚糖酶细菌的分离、鉴定和
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