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ABSTRACT:Biological degumming is an eco-friendly, efficient, high-quality and low-cost method that has become the leading bast fiber degumming technology. However, bacterial strains with short degumming cycles, high gum removal rates and small fiber damage are few. To screen high quality microbial resources with bast-fiber biological degumming function, soil samples were collected from a continuously cultivated banana plantation and then used to be enriched by ramie and kenaf materials in turn. A selective pectin-degrading medium was used to screen for excellent bacteria. A dominant bacterial strain was identified by phenotypic and genotypic characteristics, and its biological degumming effects on ramie and kenaf were verified by a comprehensive evaluation system. Results showed that seven bacterial strains secreting pectinase were obtained and the largest hydrolysis circle with a diameter ratio H/C of 2.4 was produced by the bacterial strain hn1-1, which was preliminarily identified as the Bacillus cereus by colony morphological characteristics and 16S rDNA sequence (GenBank accession number: KX013542) cluster analysis. The fiber production of ramie and kenaf degummed by B. cereus hn1-1 for 10 h were 72 % and 76 %, the residual gum rates were 4 % and 5 %, respectively. These values satisfied the textile industry requirement of < 6 % residual gum rate. Therefore, an effective biological degumming bacterium, B. cereus, was identified using a pectin-hydrolysis selective medium through a simple, economical, and time-saving method. Furthermore, the biological degumming technology by B. cereus for ramie and kenaf had a short cycle, ideal removal gum rate, and high-quality and productive fiber output.

A Gram-negative, non-motile, non-spore-forming bacterial strain designated IBFC2009(T) was isolated from soil of a bamboo plantation. The strain could grow at 11 degrees C approximately 39 degrees C, pH 6.0-9.0, and in the presence of 0 approximately 5% NaCl. Based on 16S rRNA gene sequence analysis, Strain IBFC2009(T) belonged to the genus Sphingobacterium and showed the highest sequence similarity of 94.6% (S. composti T5-12(T)) with the type strains within the genus. The major fatty acids were summed feature 3 (iso-C(15:0) 2-OH and/or C(16:1) omega7c, 34.4%), iso-C(15:0) (22.4%), C(16:0) 3-OH (15.2%), and iso-C(17:0) 3-OH (12.8%). The G+C content of the genomic DNA was 41.0 mol%. According to the phenotypic and genotypic characteristics, Strain IBFC2009(T) should represent a novel species of the genus Sphingobacterium, for which the name Sphingobacterium bambusae sp. nov. is proposed. The type strain is IBFC2009(T) (=CCTCC AB 209162(T) =KCTC 22814(T)).

Factory tests for rapid process of ramie bio-degumming by Pectobacterium sp. CXJZU-120 were conducted and evaluated comparing the processes of traditional chemical degumming and bio-chemical degumming. Results revealed that over 90% of the gum in raw ramie could be removed only with Pectobacterium sp. CXJZU-120 in 6 h. The rapid process was not only suitable for the extraction of ramie fibers from different grades of raw material and retaining the inherent morphological structures and textile properties, but also could reduce the production cost up to 20.5%, raise resource utilization by more than 50% and reduce pollution charge by more than 80% compared with the traditional chemical degumming. It is a breakthrough in the degumming of ramie and has great application potential in the extraction of herbaceous fiber materials.

Xylanases catalyze the hydrolysis of xylan, a major hemicellulose component of cell wall besides cellulose in most plant species. To extract cellulose fibers, it will be invaluable to screen for more effective xylanase-producing microorganisms. In this paper a new strategy for easy screening of xylanase-producing strains from the degumming line was presented. Using this strategy, a weak-acidic, cellulase-free xylanase from Bacillus subtilis has been isolated, purified and characterized. The xylanase showed high specific activity (36,633.4 U/mg), presented stable characteristics and can be separated and purified simply, with molecular weight 23.3 kD, pI 9.63. It reached its optimal activity at pH 5.8 and 60 °C, and retained over 80% of its maximal activity after pre-incubation at temperature 60 °C or pH 4.6 ~ 6.4. Also, a two-step purification procedure based on the combination of ultrafiltration and gel filtration chromatography was introduced and described, achieving 17-fold purification with 68.11% yield.

β-mannanase has shown compelling biological functions because of its regulatory roles in metabolism, inflammation, and oxidation. This study separated and purified the β-mannanase from Bacillus subtilis BE-91, which is a powerful hemicellulose-degrading bacterium using a “two-step” method comprising ultrafiltration and gel chromatography. The purified β-mannanase (about 28.2 kDa) showed high specific activity (79, 859.2 IU/mg). The optimum temperature and pH were 65°C and 6.0, respectively. Moreover, the enzyme was highly stable at temperatures up to 70°C and pH 4.5–7.0. The β-mannanase activity was significantly enhanced in the presence of Mn2+, Cu2+, Zn2+, Ca2+, Mg2+, and Al3+ and strongly inhibited by Ba2+ and Pb2+. K m and V max values for locust bean gum were 7.14 mg/mL and 107.5 μmol/min/mL versus 1.749 mg/mL and 33.45 µmol/min/mL for Konjac glucomannan, respectively. Therefore, β-mannanase purified by this work shows stability at high temperatures and in weakly acidic or neutral environments. Based on such data, the β-mannanase will have potential applications as a dietary supplement in treatment of inflammatory processes.

Among industrial fiber crops, jute is ranked second to cotton in terms of yield and planting area worldwide. The traditional water retting and chemical semi-degumming methods restrict the development of the jute industry. Jute fiber can be extracted from jute bast through mechanical rolling (preprocessing), culture of bacteria, soaking fermentation (liquor ratio = 10, inoculum size = 1 %, temperature = 35 °C, and time = 15 h), inactivation, washing, and drying. Pectobacterium sp. DCE-01 secretes key degumming enzymes: pectinase, mannase, and xylanase, which match well the main non-cellulosic components of jute bast. Compared with the traditional water retting degumming, the bio-degumming cycle is shortened from more than 10 days to 15 h. The proposed bio-degumming achieved higher efficiency and lower pollution than water retting and chemical semi-degumming.

Pectinase plays a crucial role in ramie bio-degumming. A pectate lyase gene ( pel4J4) from the high-efficiency degumming bacteria Dickeya dadantii DCE-01 of bast fibers was cloned and connected to pET28a, and then the recombinant plasmid was successfully transformed into Escherichia coli BL21(DE3). The pectate lyase (Pel4J4) induced was purified by ultrafiltration and Sephadex G-100 gel chromatography. The enzymatic properties of Pel4J4 were studied in detail. pel4J4 (GenBank accession number: KC900167) had a sequence length of 1179 bp, encoding 392 amino acids. The extracellular pectate lyase activity of pET28a- pel-BL was up to 204.4 IU/mL. The optimal temperature and pH of the purified Pel4J4 were 55℃ and 8.5, respectively. The stable temperature and pH of Pel4J4 activity were 45℃ and 8.5–10.0, respectively. The catalytic activity is Ca2+ dependent and promoted by 1 mmol/L Zn2+, Fe3+, Ca2+, and NH4+, but seriously inhibited by Cu2+ and Pb2+. The optimal substrate is citrus pectin with more than 85% esterification. The heat-resistant alkaline Pel4J4 could strongly degrade natural ramie pectin, indicating a promising application prospect in ramie bio-degumming.