Page 227 - 《华中农业大学学报(自然科学版)》2022年第5期
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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
                                                                    characterization of a novel chitosanase from Streptomyces al⁃
               高,培养瓶底部堆积,减少固体基质内气体含量和气
                                                                    bolongus ATCC 27414[J].Food chemistry,2019,286:696-702.
               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
                                                                    及酶学性质分析[J]. 湖北农业科学,2016,55(7):1778-1781.LI
                     [14]
               U/mL) 、木霉属菌株 T. harzianum(1.40 U/mL)、                J Y,WANG H M,ZU G R.Study on the inducing conditions and
               T. koningii(1.20 U/mL)、T. viride(1.02 U/mL)和 T.      enzyme characteristics of a strain marine fungal MF-08 producing
                                                                    chitosanase[J].Hubei agricultural sciences,2016,55(7):1778-
               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.
                 [5]
               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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