Production Of Oligosaccharides Research Articles

De-Oiled Citrus Peels as Feedstock for the Production of Pectin Oligosaccharides and Its Effect on Lactobacillus fermentum, Probiotic Source.

Following the extraction of essential oil, citrus (Mousambi, Kinnow, and Orange) peel wastes were used to produce pectin. The yield of essential oil and pectin was maximum in orange. Pectin was characterized by Fourier-transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) spectroscopy. The degree of esterification (DE) and methoxyl content (MC) was maximum in orange whereas, the equivalent weight was maximum in Mousambi. A significant increase (61.8%) in the Lactobacillus fermentum population was observed with pectin as compared with sugar. Three sources followed the Orange > Kinnow > Mousambi trend as a prebiotic source. It was attributed to higher DE as well as higher MC. Enhancement in the bacterial population was in the range of 79.16–87.50%. The present work confirms the potential of pectin as a probiotic source for the enhancement of the bacterial population. Thus, it has a large scope for use in the food industry targeting a circular economy.

Valorization of corn cobs for xylanase production by Aspergillus flavus AW1 and its application in the production of antioxidant oligosaccharides and removal of food stain

Xylanases are hemicellulolytic enzymes specifically catalyze xylan hydrolysis that constitutes the most abundant polysaccharide in the hemicellulose fraction of the lignocellulosic biomasses. The effective conversion of these biomasses to valuable commodities required the use of economic effective biocatalysts . In this study, xylanase production was performed by corn cobs fermentation using Aspergillus flavus AW1 in which the highest productivity was 356.36 U/g dry substrate. The partially purified enzyme was aflatoxin-free as estimated by high performed liquid chromatography . The enzyme was optimally active over a wide pH range from 5.5 to 7.5. Additionally, it was optimally active at temperature of 55 °C and at 60 °C it possessed 95% of its maximum activity. The enzyme half-lives were estimated as 83.81 and 44.69 min at 55 and 60 ° C, respectively. The V max and K m for the purified enzyme fraction were 250 U/mg protein/min and 5.55 mg/mL, respectively. Moreover, beech wood xylan hydrolysis as well as the hydrolysis of the hemicellulosic fraction of corn cobs was examined by applying the produced enzyme. The obtained results indicated hydrolysis percentage values of 39 and 19% for beech wood xylan and the hemicellulosic fraction of corn cobs, respectively with the release of a mixture of xylooligosaccharides confirmed by thin layer and high performed liquid chromatography analysis. The antioxidant activities for the dried oligosaccharide mixtures were estimated by scavenging activity of 1,1-diphenyl-2-picrylhydrazyl free radical. Finally, the efficiency of the purified enzyme fraction in the removal of mango stain was investigated in comparison to the commercial detergent. • Xylanase was produced by solid state fermentation of corn cobs using Aspergillus flavus AW1. • The partially purified enzyme was thermo-active and possessed high affinity toward beech wood xylan. • The enzyme was applied for the removal of food stain and production of xylooligosaccharides. • The produced xylooligosaccharides possessed antioxidant activity.

Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis.

Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70°C for 1 h. The Km and Vmax of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.

Galacto-Oligosaccharide (GOS) Synthesis during Enzymatic Lactose-Free Milk Production: State of the Art and Emerging Opportunities

Much attention has recently been paid to β-Galactosidases (β-D-galactoside galactohidrolase; EC 3.2.1.23), commonly known as lactases, due to the lactose intolerance of the human population and the importance of dairy products in the human diet. This enzyme, produced by microorganisms, is being used in the dairy industry for hydrolyzing the lactose found in milk to produce lactose-free milk (LFM). Conventionally, β-galactosidases catalyze the hydrolysis of lactose to produce glucose and galactose in LFM; however, they can also catalyze transgalactosylation reactions that produce a wide range of galactooligosaccharides (GOS), which are functional prebiotic molecules that confer health benefits to human health. In this field, different works aims to identify novel microbial sources of β-galactosidase for removing lactose from milk with the relative GOS production. Lactase extracted from thermophilic microorganisms seems to be more suitable for the transgalactosylation process at relatively high temperatures, as it inhibits microbial contamination. Different immobilization methods, such as adsorption, covalent attachment, chemical aggregation, entrapment and micro-encapsulation, have been used to synthesize lactose-derived oligosaccharides with immobilized β-galactosidases. In this mini-review, particular emphasis has been given to the immobilization techniques and bioreactor configurations developed for GOS synthesis in milk, in order to provide a more detailed overview of the biocatalytic production of milk oligosaccharides at industrial level.

Mechanistic insights into consumption of the food additive xanthan gum by the human gut microbiota.

Processed foods often include food additives such as xanthan gum, a complex polysaccharide with unique rheological properties, that has established widespread use as a stabilizer and thickening agent. Xanthan gum's chemical structure is distinct from those of host and dietary polysaccharides that are more commonly expected to transit the gastrointestinal tract, and little is known about its direct interaction with the gut microbiota, which plays a central role in digestion of other dietary fibre polysaccharides. Here we show that the ability to digest xanthan gum is common in human gut microbiomes from industrialized countries and appears contingent on a single uncultured bacterium in the family Ruminococcaceae. Our data reveal that this primary degrader cleaves the xanthan gum backbone before processing the released oligosaccharides using additional enzymes. Some individuals harbour Bacteroides intestinalis that is incapable of consuming polymeric xanthan gum but grows on oligosaccharide products generated by the Ruminococcaceae. Feeding xanthan gum to germfree mice colonized with a human microbiota containing the uncultured Ruminococcaceae supports the idea that the additive xanthan gum can drive expansion of the primary degrader Ruminococcaceae, along with exogenously introduced B. intestinalis. Our work demonstrates the existence of a potential xanthan gum food chain involving at least two members of different phyla of gut bacteria and provides an initial framework for understanding how widespread consumption of a recently introduced food additive influences human microbiomes.

Development of an innovative macroalgae biorefinery: Oligosaccharides as pivotal compounds

Macroalgae have significant advantages over land-living biomass resources and are promising pivotal feedstocks for the onset of the blue bioeconomy. Among these, Ulva lactuca has demonstrated a high potential due to its wide distribution and high productivity.In this work, a detailed chemical characterization of U. lactuca enabled the identification of polysaccharides as the main macromolecular component of the organic fraction. They present a high diversity of sugar constituents and hence can be a relevant source of added-value oligosaccharides for the food/feed industries.Four processes, with increasing operational temperatures, were compared for the selective production of oligosaccharides: Conventional Soxhlet Extraction, Accelerated Solvent Extraction, Hydrothermal treatment (HT) and Dilute Acid Hydrolysis (DAH). All processes presented high oligosaccharide/monosaccharide ratios, with HT and DAH exhibiting the highest oligosaccharides yields (10.6 and 16.6 g/100 g initial biomass, respectively). These oligosaccharides were obtained under milder, more economic conditions than the reported for lignocellulosic (land) plants and can represent an important added-value income of the algae biorefineries and thus contribute to their economic sustainability.

Toward Understanding the Alginate Catabolism in Microbulbifer sp. ALW1 by Proteomics Profiling.

The bacterial strain of Microbulbifer sp. ALW1 has demonstrated visible ability of degrading the cell wall of Laminaria japonica, and biochemical characterization has been performed on some individual enzymes to elucidate its genetic basis. However, it still remains elusive how strain ALW1 successfully breaks down the major cell wall component alginate polysaccharide and colonizes on its marine host. In this study, a mass spectrometry-based quantitative analysis of the extracellular and intracellular proteomes was introduced to elucidate the alginate degradation pathway in ALW1 strain. Mass spectrometry and biochemical assays indicated that strain ALW1 could effectively degrade alginate polysaccharide into disaccharides and trisaccharides within 12 h. Proteome analysis identified 156 and 1,047 proteins exclusively localized in extracellular and intracellular compartments, respectively, with 1,086 protein identities of dual localization. Functional annotation of the identified proteins suggested the involvement of diverse catalytic enzymes and non-catalytic molecules for the cleavage and metabolism of alginate polysaccharide. A simplified pathway was constructed to demonstrate the extracellular digestion, active transport, and intracellular conversion of alginate polysaccharide and its fragmented oligosaccharides, casting a picture of genetic loci controlling alginate catabolism by ALW1 strain. This study aims to provide a guide for utilization and genetic manipulation of the bacterial strain ALW1 for efficient alginate oligosaccharides production by fermentation.

Emerging trends of microbial technology for the production of oligosaccharides from biowaste and their potential application as prebiotic

Oligosaccharides are the sugars made up of 3–10 saccharides units and one of the classes of prebiotics obtained from various biowastes. These biowastes could include rice straw, husk, spent coffee grounds, sugarcane bagasse, spent tea leaves, fruits and vegetables peel, corn stalk, corn stover, deoiled meals and brewer's spent grains etc., which can be used as a resource for oligosaccharides production. This review aims to provide a comprehensive overview of the suitability of different biowaste resources for oligosaccharide production followed by critical analysis of the recent updates and production methods. The review also discusses the tremendous prebiotic potential of oligosaccharides in food applications with prospects for further advancements in the field.

Heterogenously‐expressed chitosanase combining a green ball milling method for enzymatic degradation

AbstractChitooligosaccharides (COSs) and d‐glucosamine (GlcN), the most valuable biomolecules due to their various physiological functions, can be available inexpensively from chitosan by bio‐enzymatic degradation, in which chitosanases play a key role. On the other hand, pretreatment of chitosan is the necessary procedure in the bioproduction by enzymatic hydrolysis. Therefore, the green and efficient pretreatment strategy as well as superior enzyme activity are the most critical aspects in scaled up production. This study investigated environmentally friendly and efficient enzymatic hydrolysis of powdery chitosan (PC) pretreated by ball‐milling. First, heterologous expression yielded OUC‐CsnPT, a glycoside hydrolase family 46 chitosanase with a predominant specific activity of 5346.56 U/mg at pH 6.0°C and 45°C. The endo‐type chitosanase OUC‐CsnPT hydrolyzed chitosan to produce GlcN and chitobiose [(GlcN)2]. Then, ball‐milling was employed to pretreat PC to promote chitosanase OUC‐CsnPT hydrolysis, and the effectiveness was characterized by scanning electron microscope, X‐ray diffraction, and FTIR‐attenuated total reflection. Through the development of the novel route, the yield of COSs and GlcN for ball‐milling powdery chitosan increased by 10.8‐folds with a concentration of 14.5 μmol/ml. Hence, this technique presents a promising strategy suited to be a straightforward and environmentally friendly option for oligosaccharides production, broadening the theoretical basis for PC biodegradation.

Biochemical Properties of a New Polysaccharide Lyase Family 25 Ulvan Lyase TsUly25B from Marine Bacterium Thalassomonas sp. LD5.

Marine macroalgae, contributing much to the bioeconomy, have inspired tremendous attention as sustainable raw materials. Ulvan, as one of the main structural components of green algae cell walls, can be degraded by ulvan lyase through the β-elimination mechanism to obtain oligosaccharides exhibiting several good physiological activities. Only a few ulvan lyases have been characterized until now. This thesis explores the properties of a new polysaccharide lyase family 25 ulvan lyase TsUly25B from the marine bacterium Thalassomonas sp. LD5. Its protein molecular weight was 54.54 KDa, and it was most active under the conditions of 60 °C and pH 9.0. The Km and kcat values were 1.01 ± 0.05 mg/mL and 10.52 ± 0.28 s−1, respectively. TsUly25B was salt-tolerant and NaCl can significantly improve its thermal stability. Over 80% of activity can be preserved after being incubated at 30 °C for two days when the concentration of NaCl in the solution is above 1 M, while 60% can be preserved after incubation at 40 °C for 10 h with 2 M NaCl. TsUly25B adopted an endolytic manner to degrade ulvan polysaccharides, and the main end-products were unsaturated ulvan disaccharides and tetrasaccharides. In conclusion, our research enriches the ulvan lyase library and advances the utilization of ulvan lyases in further fundamental research as well as ulvan oligosaccharides production.

Biochemical Characterization and Cold-Adaption Mechanism of a PL-17 Family Alginate Lyase Aly23 from Marine Bacterium Pseudoalteromonas sp. ASY5 and Its Application for Oligosaccharides Production.

As an important enzyme involved in the marine carbon cycle, alginate lyase has received extensive attention because of its excellent degradation ability on brown algae, which is widely utilized for alginate oligosaccharide preparation or bioethanol production. In comparison with endo-type alginate lyases (PL-5, PL-7, and PL-18 families), limited studies have focused on PL-17 family alginate lyases, especially for those with special characteristics. In this study, a novel PL-17 family alginate lyase, Aly23, was identified and cloned from the marine bacterium Pseudoalteromonas carrageenovora ASY5. Aly23 exhibited maximum activity at 35 °C and retained 48.93% of its highest activity at 4 °C, representing an excellent cold-adaptation property. Comparative molecular dynamics analysis was implemented to explore the structural basis for the cold-adaptation property of Aly23. Aly23 had a high substrate preference for poly β-D-mannuronate and exhibited both endolytic and exolytic activities; its hydrolysis reaction mainly produced monosaccharides, disaccharides, and trisaccharides. Furthermore, the enzymatic hydrolyzed oligosaccharides displayed good antioxidant activities to reduce ferric and scavenge radicals, such as hydroxyl, ABTS+, and DPPH. Our work demonstrated that Aly23 is a promising cold-adapted biocatalyst for the preparation of natural antioxidants from brown algae.

A Comparison of Mother's Milk and the Neonatal Urine Metabolome: A Unique Fingerprinting for Different Nutritional Phenotypes.

The ability of metabolomics to provide a snapshot of an individual’s metabolic state makes it a very useful technique in neonatology for investigating the complex relationship between nutrition and the state of health of the newborn. Through an 1H-NMR metabolomics analysis, we aimed to investigate the metabolic profile of newborns by analyzing both urine and milk samples in relation to the birth weight of neonates classified as AGA (adequate for the gestational age, n = 51), IUGR (intrauterine growth restriction, n = 14), and LGA (large for gestational age, n = 15). Samples were collected at 7 ± 2 days after delivery. Of these infants, 42 were exclusively breastfed, while 38 received mixed feeding with a variable amount of commercial infant formula (less than 40%) in addition to breast milk. We observed a urinary spectral pattern for oligosaccharides very close to that of the corresponding mother’s milk in the case of exclusively breastfed infants, thus mirroring the maternal phenotype. The absence of this good match between the infant urine and human milk spectra in the case of mixed-fed infants could be reasonably ascribed to the use of a variable amount of commercial infant formulas (under 40%) added to breast milk. Furthermore, our findings did not evidence any significant differences in the spectral profiles in terms of the neonatal customize centile, i.e., AGA (adequate for gestational age), LGA (large for gestational age), or IGUR (intrauterine growth restriction). It is reasonable to assume that maternal human milk oligosaccharide (HMO) production is not or is only minimally influenced by the fetal growth conditions for unknown reasons. This hypothesis may be supported by our metabolomics-based results, confirming once again the importance of this approach in the neonatal field.

C-Terminal Bacterial Immunoglobulin-like Domain of κ-Carrageenase Serves as a Multifunctional Module to Promote κ-Carrageenan Hydrolysis.

κ-Carrageenase is an important component for κ-carrageenan oligosaccharide production. Generally, noncatalytic domains are appended to carbohydrate-active domains and potentiate catalytic activity. However, studies devoted to κ-carrageenase are relatively few. Here, a C-terminal bacterial immunoglobulin-like domain (Big_2) was identified in κ-carrageenase (PpCgk) from Pseudoalteromonas porphyrae. Biochemical characterization of native PpCgk and its two truncations, PpCgkCD (catalytic domain) and PpBig_2 (Big_2 domain), revealed that the specific activity, catalytic efficiency (kcat/Km(app)), specific κ-carrageenan-binding capacity, and thermostability of PpCgk were significantly higher than those of PpCgkCD, suggesting that the noncatalytic PpBig_2 domain is a multifunctional module and essential for maintaining the activity and thermostability of PpCgk. Furthermore, it was found that the mode of action of PpCgk was more processive on both the dissolved and gelled substrates than that of PpCgkCD, indicating that PpBig_2 contributes to the processivity of PpCgk. Interestingly, PpBig_2 can be used as an independent module to enhance the hydrolysis of κ-carrageenan through its disruptive function. In addition, sequence analysis suggests that Big_2 domains are highly conserved in bacterial κ-carrageenases, implying the universality of their noncatalytic functions. These findings reveal the multifunctional role of the noncatalytic PpBig_2 and will guide future functional analyses and biotechnology applications of Big_2 domains.

Marine microbial enzymes for the production of algal oligosaccharides and its bioactive potential for application as nutritional supplements.

Marine macroalgae have a very high carbohydrate content due to complex algal polysaccharides (APS) like agar, alginate, and ulvan in their cell wall. Despite numerous reports on their biomedical properties, their hydrocolloid nature limits their applications. Algal oligosaccharides (AOS), which are hydrolyzed forms of complex APS, are gaining importance due to their low molecular weight, biocompatibility, bioactivities, safety, and solubility in water that makes it a lucrative alternative. The AOS produced through enzymatic hydrolysis using microbial enzymes have far-reaching applications because of its stereospecific nature. Identification and characterization of novel microorganisms producing APS hydrolyzing enzymes are the major bottlenecks for the efficient production of AOS. This review will discuss the marine microbial enzymes identified for AOS production and the bioactive potential of enzymatically produced AOS. This can improve our understanding of the biotechnological potential of microbial enzymes for the production of AOS and facilitate the sustainable utilization of algal biomass. Enzymatically produced AOS are shown to have bioactivities such as antioxidant, antiglycemic, prebiotic, immunomodulation, antiobesity or antihypercholesterolemia, anti-inflammatory, anticancer, and antimicrobial activity. The myriad of health benefits provided by the AOS is the need of the hour as there is an alarming increase in physiological disorders among a wide range of the global population.

Oligosaccharide production from preserved yuzu juice using Lactobacillus sakei NY 518 and its prebiotic function

This study focused on the physiochemical characteristics of the oligosaccharide produced in preserved yuzu by NY 518 and its functions based on the intestinal health through antimicrobial activities. NY 518 was identified as Lactobacillus sakei. The oligosaccharide produced by L. sakei NY 518 was gluco-oligosaccharide. The oligosaccharide from preserved yuzu (OY) was found to have lower degree of polymerization than the oligosaccharide from sucrose (OS) owing to the presence of tri- to nona-saccharides. These oligosaccharides had lower pH and increased acidity, soluble solids, and viscosity than their substrates. OY had 67% α-glucosidase inhibition activity of OS and higher antibacterial effects on food-poisoning bacteria than OS. These results indicate that the oligosaccharide produced by L. sakei NY 518 in sugar-preserved foods exhibits beneficial functions (antidiabetic and antibacterial activities) and could be a good substitute for sugar.

Production, characterization, and prebiotic activity of oligosaccharides from konjac glucomannan by Bacillus amyloliquefaciens WX-1

Konjac glucomannan (KGM) oligosaccharides from KGM is a potentially and valuable prebiotics. A KGM degrading strain was screened and identified as Bacillus amyloliquefaciens WX-1. B. amyloliquefaciens WX-1 was shown to efficiently depolymerize KGM and produce a broad range of small molecule oligosaccharides. The strain efficiently processed KGM with 752.9 mg g−1 yields of KGM oligosaccharides (KGOS) at optimal conditions. The KGOS were characterized using TLC, MALDI-TOF-MS and NMR spectroscopy. The result of KGOS composition revealed that it contained a series of oligosaccharides with a degree of polymerization of 2–14 and the disaccharide structures of MM, MG and GM were assumed. The prebiotic function of KGOS to the growth of Lactobacillus was evaluated. This study showed that B. amyloliquefaciens WX-1 provided a basis for the further development as biocatalyst for direct conversion of polysaccharides derived from energy crops as well as agricultural residues to functional prebiotics.

Combination of Autohydrolysis and Catalytic Hydrolysis of Biomass for the Production of Hemicellulose Oligosaccharides and Sugars

Three different types of biomass sourced from forestry waste (eucalyptus residues), agricultural waste (wheat straw), and energy crop (miscanthus) were used as starting materials to produce hemicellulosic sugars, furans (furfural and hydroxymethylfurfural), and oligosaccharides. A two-step hybrid process was implemented; biomass was first autohydrolysed without any additive to extract hemicelluloses and dissolve it in water. Then, the hydrolysate was treated with a solid acid catalyst, TiO2-WOx, in order to achieve hydrolysis and produce monomeric sugars and furans. This article investigates the role of the biomass type, autohydrolysis experimental conditions, polymerisation degree and composition of hemicelluloses on the performance of the process coupling autohydrolysis and catalytic hydrolysis. The highest global yields of both oligosaccharides and monomeric sugars were obtained from Eucalyptus (37% and 18%, respectively).

Production of prebiotic gellan oligosaccharides based on the irradiation treatment and acid hydrolysis of gellan gum

Biologically active gellan oligosaccharides (GOSs), newly found plant elicitors and biostimulants, are produced from the hydrolysis of gellan gum. Traditional hydrolysis with concentrated acid suffers from the problems of high pollution and low functional oligosaccharide yield because the process is difficult to control. Irradiation (60Co γ-ray) with a dosage ranging from 0 kGy to 175 kGy was used to degrade gellan gum efficiently and cleanly into low molecular weight (Mw) gellan with an average Mw ranging from 449,119 Da to 72,903 Da. The low Mw gellan irradiated at 70 kGy was further hydrolyzed with low concentration acid (0.5 mol/L HCl) to produce GOSs with DPs mainly 4 and 8, indicating that the Rha-β-(1 → 3)-Glc bonds in gellan gum were easily cut to produce residues with tetrasaccharide repeat subunits. Besides antioxidant activity, GOSs were also proved with prebiotic activity by in vitro fecal fermentation in a self-designed bionic intestinal reactor.

Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides.

Chitosanase plays an important role in enzymatic production of chitosan oligosaccharides (COSs). The present study describes the gene cloning and high-level expression of a high-efficiency chitosanase from Bacillus mojavensis SY1 (CsnBm). The gene encoding CsnBm was obtained by homologous cloning, ligated to pPICZαA, and transformed into Pichia pastoris X33. A recombinant strain designated X33-C3 with the highest activity was isolated from 120 recombinant colonies. The maximum activity and total protein concentration of recombinant strain X33-C3 were 6,052 U/ml and 3.75 g/l, respectively, which were obtained in fed-batch cultivation in a 50-l bioreactor. The optimal temperature and pH of purified CsnBm were 55°C and 5.5, respectively. Meanwhile, CsnBm was stable from pH 4.0 to 9.0 and 40 to 55°C. The purified CsnBm exhibited high activity toward colloidal chitosan with degrees of deacetylation from 85 to 95%. Furthermore, CsnBm exhibited high efficiency to hydrolyze different concentration of colloidal chitosan to produce COSs. The result of this study not only identifies a high-efficiency chitosanase for preparation of COSs, but also casts some insight into the high-level production of chitosanase in heterologous systems.

Self-assembled κ-carrageenase-inorganic hybrid nanoflowers exerting high catalytic efficiency with stable and recyclable properties

κ-Carrageenan oligosaccharides from κ-carrageenan hydrolysis are important biochemicals with more bioactivity. Enzyme engineering plays a key role in improving κ-carrageenase catalytic efficiency for production of κ-carrageenan oligosaccharides. Effect of metal ions on enzyme activity, especially stability and efficiency, is main factor in catalytic process, but metal ions addition leads to gelation of κ-carrageenan solution. In this study, molecular dynamics simulation was used to explore the interaction between κ-carrageenase CgkPZ and Ca2+, and Ca2+ bonded to D164 and E167 in the catalytic center resulting in the catalytic efficiency increase. Circular dichroism analysis indicated that the secondary structure of κ-carrageenase could change in the presence of Ca2+. Therefore, a novel self-assembly κ-carrageenase-inorganic hybrid nanoflowers CaNF@CgkPZ was synthesized and systematically characterized. The catalytic efficiency (kcat/Km) of CaNF@CgkPZ was 382.1 mL·mg−1·s−1, increased by 292% compared with free κ-carrageenase. Notably, the enzyme activity of CaNF@CgkPZ was not reduced significantly after 19 cycles use, and 70–100% relative activity was still retained when stored at 4–25 ℃ for 15 days. This work provides an efficient approach for κ-carrageenase immobilization with good storage stability, reusability and enhanced catalytic efficiency, which is of great significance in practical applications.